From owner-proteins@net.bio.net Thu Feb 01 22:00:00 1996 Path: biosci!mani.unet.umn.edu!npv From: npv@mani.unet.umn.edu (Nora Plesofsky-Vig) Newsgroups: bionet.molbio.proteins Subject: serial immunoprecipitation Date: 2 Feb 1996 14:40:07 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: <9602022024.AA02236@mani.cbs.umn.edu> Reply-To: nora@biosci.cbs.umn.edu NNTP-Posting-Host: net.bio.net Does anyone have experience with co-immunoprecipitating proteins, solubilizing the proteins, and reacting them with another antibody? We have tried solubilizing an immunoprecipitate with 4% SDS, diluting it to 0.1% SDS for a subsequent antibody reaction, but this did not produce positive results. We will now try a final concentration of 0.05% SDS, as well as different concentrations of urea (using either 8 M or 4 M for the initial solubilization). If you have any experience with this approach or other reagents that have worked, I would really appreciate learning of them. Thanks very much. Nora Plesofsky-Vig nora@biosci.cbs.umn.edu From owner-proteins@net.bio.net Thu Feb 01 22:00:00 1996 Path: biosci!daresbury!bioftp.unibas.ch!infobiogen.fr!jussieu.fr!univ-lyon1.fr!in2p3.fr!swidir.switch.ch!cisun2000.unil.ch!news From: Henry Hugues Newsgroups: bionet.molbio.proteins Subject: Re: Chemiluminescence Date: 2 Feb 1996 14:21:45 GMT Organization: CHUV / LCC-BH-18 Lines: 6 Message-ID: <4et6lp$3en@cisun2000.unil.ch> References: <4dd78e$3hd@studium.student.umu.se> <4ej7an$grn@sifon.cc.mcgill.ca> NNTP-Posting-Host: mac1bh18228.hospvd.ch Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; PPC) To: surglab1@is.rvh.mcgill.ca X-URL: news:4ej7an$grn@sifon.cc.mcgill.ca We have obtained our best results using the ECL kit (Amersham) with PVDF membrane. The best blocking reagent (better than BSA or Gelatine) we found is BLOTTO (powered defat. milk) 2% or 5% (w/w) in PBS. Good luck From owner-proteins@net.bio.net Thu Feb 01 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: smittola@aol.com (SMITTOLA) Newsgroups: bionet.molbio.proteins Subject: Peptide Metal Complexes? Date: 2 Feb 1996 02:55:38 -0500 Organization: America Online, Inc. (1-800-827-6364) Lines: 12 Sender: root@newsbf02.news.aol.com Message-ID: <4esg1q$d6q@newsbf02.news.aol.com> Reply-To: smittola@aol.com (SMITTOLA) NNTP-Posting-Host: newsbf02.mail.aol.com We are researching new fungicide formulations. We would like to try complexing His-Gly-Lys or Gly-Gly-His with Cu. Can anyone with experience in this area give me an idea of the ratio of peptide to Cu to make a good solid peptide-Cu complex. The custom peptides are quite expensive, so of coarse we want to get by with as little as possible per given amount of Cu. Thanks, Steve BRF Inc. Colorado From owner-proteins@net.bio.net Thu Feb 01 22:00:00 1996 Path: biosci!bloom-beacon.mit.edu!senator-bedfellow.mit.edu!knut From: knut@iti.org (Knut Langsetmo) Newsgroups: bionet.molbio.proteins Subject: Re: Press Release: A new Gel Imaging software package Date: 2 Feb 1996 15:23:46 GMT Organization: MIT Department of Biology Lines: 10 Message-ID: <4etaa2$9au@senator-bedfellow.MIT.EDU> References: <4ejdnb$650@Vir.com> NNTP-Posting-Host: rosa.mit.edu In article <4ejdnb$650@Vir.com> comcul@montreal.com writes: >Press Release: A new Gel Imaging software package ... >Although pricing for MATRIX has not yet been determined, the package >is expected to be around $1,500 (USD). It is the first Windows-based ... NIH Image is free -- --knut knut@rosa.mit.edu From owner-proteins@net.bio.net Thu Feb 01 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.wwa.com!news From: nc26@wwa.com Newsgroups: bionet.molbio.proteins Subject: Digestion of Core Protein Date: 3 Feb 1996 00:58:11 GMT Organization: WorldWide Access (tm) - Chicagoland Internet Services (http://www.wwa.com) Lines: 4 Message-ID: <4eubv3$dgr@kirin.wwa.com> NNTP-Posting-Host: vh3-024.wwa.com X-Newsreader: SPRY News 3.03 (SPRY, Inc.) Had anyone had any experiences with enzymatic and/or chemical cleavage of core protein? I've been attempting to digest HBV core protein with tryspin in 1M urea with little success. My digestion have resulted in fragments that are only partial cleave. I've gone as high as a 1:3 protein:enzyme ratio without any effect. I would appreciate any help, especially from anyone who had previous experiences with digestion of core protein. Thanks in advance. From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Path: biosci!daresbury!bioftp.unibas.ch!infobiogen.fr!jussieu.fr!univ-lyon1.fr!in2p3.fr!swidir.switch.ch!swsbe6.switch.ch!scsing.switch.ch!news.belwue.de!fu-berlin.de!zrz.TU-Berlin.DE!cs.tu-berlin.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins Subject: Re: staining of phosphoproteins Date: 2 Feb 1996 16:06:39 GMT Organization: University of Wuerzburg, Germany Lines: 16 Message-ID: <4etcqf$i9c@winx03.informatik.uni-wuerzburg.de> References: <4eqp2a$6nq@n.ruf.uni-freiburg.de> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Jens Krieger (krieger@mibm1.ruf.uni-freiburg.de) wrote: > Is there anyone "outside" who will know a recipe for staining > phosphoproteins on acrylamidgels, or could give me any hint? Silver? Copper chloride (sold by BioRad, claimed to be more sensitive than Coomassie)? The fluorescent dyes sold by Molecular Probes? (I have only experience with silver staining, and although it is very sensitive it is probably not what you want when you have to stain a lot of gels.) --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Path: biosci!lhc.nlm.nih.gov!darwin.sura.net!maze.dpo.uab.edu!usenet From: "N.Ambalavanan MD" Newsgroups: bionet.molbio.proteins Subject: Cell lysates for bFGF- How? Date: Sat, 03 Feb 1996 09:23:41 -0600 Organization: UAB Lines: 8 Message-ID: <31137DFD.50E6@uabdpo.dpo.uab.edu> NNTP-Posting-Host: tty11.maze.ppp.uab.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b6a (Win95; I) Hi! I plan to estimate bFGF production by smooth muscle cells in vitro grown in 75cm2 flasks. I want to estimate the bFGF in the conditioned media as well as in the cell lysates by ELISA. Does anyone have a protocol for producing cell lysates suitable for ELISA ? Please email me a protocol or tell me where I can find a good one. Thanks! Ambal MD UAB From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!dziuxsolim.rutgers.edu!er6.rutgers.edu!not-for-mail From: edso@eden.rutgers.edu (Edward H. So) Newsgroups: bionet.molbio.proteins Subject: Need help with Km values Date: 3 Feb 1996 08:40:02 -0500 Organization: Rutgers University Lines: 8 Message-ID: <4evoji$kd@er6.rutgers.edu> NNTP-Posting-Host: er6.rutgers.edu Hi, Does anyone have the Km values for catalase and glutathione peroxidase handy or know where I can find those values?? Really appreciate it if anybody can help me out. Thanks in advance. -Edward edso@eden.rutgers.edu From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!EU.net!Portugal.EU.net!news.rccn.net!scsing.switch.ch!news.belwue.de!fu-berlin.de!zrz.TU-Berlin.DE!cs.tu-berlin.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins Subject: Re: WHO KNOWS THE RATE OF TRANSLATION??!! (AA/min) Date: 1 Feb 1996 08:57:12 GMT Organization: University of Wuerzburg, Germany Lines: 11 Message-ID: <4epv98$jim@winx03.informatik.uni-wuerzburg.de> References: <4e5hga$sis@rc1.vub.ac.be> <1996Jan31.122553@ebi.ac.uk> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] higgins@ebi.ac.uk wrote: > (I have no idea if there is a Genes V) Yes, there is. (However, I liked IV better :-) --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!usenet.seri.re.kr!xpat.postech.ac.kr!usenet From: kim myung jong Newsgroups: bionet.molbio.proteins Subject: immunoprecipitation--help Date: Sat, 03 Feb 1996 21:42:21 -0800 Organization: Lab of SignalTransduction ,Dep.of Life Science, POSTECH Lines: 13 Message-ID: <3114473D.6359@vision.postech.ac.kr> Reply-To: Lab, of, SignalTransduction, Dep.of, Life, Science, POSTECH, San31, HyoJaDong, PoHang, South, Korea NNTP-Posting-Host: hybrid.postech.ac.kr Mime-Version: 1.0 Content-Type: text/plain; charset=euc-kr Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b5 (Macintosh; I; PPC) Hi! everyone. I am graduate student in south korea. I want to know tip that can reduce non-specific association of cell extract to protein A agarose in immunoprecipitation experiment. I routinly add antibody solution to pre-cleared mammalian cell extract and then add protein A agarose. In my opinion, nonspecific binding is due to washing buffer. My washing buffer contain 1% Triton X-100 and 180mM NaCl. Please, give me solution. Thanks in advance. My email address is pgs@vision.postech.ac.kr Myung Jong Kim From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!rutgers!umdnj!njmsa!upperman From: upperman@njmsa.UMDNJ.EDU (Jeffrey Upperman) Subject: Tissue protein isolation? Message-ID: Summary: tissue protein isolation? Keywords: tissue protein isolation? Sender: news@umdnj.edu () Organization: Univ. of Medicine and Dentistry of NJ Date: Thu, 1 Feb 1996 09:17:09 GMT Lines: 11 does anyone have a protocol or reference for tissue protein isolation? be gentle i am new to the protein world... -- Jeffrey S. Upperman,MD Internet: upperman@umdnj.edu Research Fellow Voice: (201) 982-4474 Dept. Anatomy, Cell Biology & Injury Fax: (201) 982-6803 Dept. of Surgery Beeper: (201) 708-4150 New Jersey Medical School Office: (201) 982-5045 185 S. Orange Ave., G606 Lab: (201) 982-5404 Newark, NJ 07103 From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Path: biosci!GAMMA.IS.TCU.EDU!XZHENG From: XZHENG@GAMMA.IS.TCU.EDU Newsgroups: bionet.molbio.proteins Subject: need help on peptide synthesizer Date: 3 Feb 1996 13:08:17 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 59 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I0S4DU5OF600DZMP@GAMMA.IS.TCU.EDU> NNTP-Posting-Host: net.bio.net From: IN%"postmaster@GAMMA.IS.TCU.EDU" "PMDF Mail Server" 3-FEB-1996 15:01:56.36 To: IN%"postmaster@GAMMA.IS.TCU.EDU", IN%"XZHENG@GAMMA.IS.TCU.EDU" CC: Subj: Undeliverable mail: SMTP delivery failure Return-path: <> Received: from GAMMA.IS.TCU.EDU by GAMMA.IS.TCU.EDU (PMDF V5.0-5 #9380) id <01I0S48VTVKW00E1ZR@GAMMA.IS.TCU.EDU>; Sat, 03 Feb 1996 15:01:48 -0600 (CST) Date: Sat, 03 Feb 1996 15:01:48 -0600 (CST) From: PMDF Mail Server Subject: Undeliverable mail: SMTP delivery failure To: postmaster@GAMMA.IS.TCU.EDU, XZHENG@GAMMA.IS.TCU.EDU Message-id: <01I0S48XGGGI00E1ZR@GAMMA.IS.TCU.EDU> MIME-version: 1.0 Content-type: MULTIPART/MIXED; BOUNDARY="Boundary (ID 01MLeX23CE1DmndbOqTJAA)" --Boundary (ID 01MLeX23CE1DmndbOqTJAA) Content-type: TEXT/PLAIN; CHARSET=US-ASCII The message could not be delivered to: Addressee: methods_and_reagents@net.bio.net : Remote system's reason for rejecting: ... User unknown --Boundary (ID 01MLeX23CE1DmndbOqTJAA) Content-type: MESSAGE/RFC822 Received: from GAMMA.IS.TCU.EDU by GAMMA.IS.TCU.EDU (PMDF V5.0-5 #9380) id <01I0S48DFFGG00DZMP@GAMMA.IS.TCU.EDU> for methods_and_reagents@net.bio.net; Sat, 03 Feb 1996 15:01:44 -0600 (CST) Date: Sat, 03 Feb 1996 15:01:44 -0600 (CST) From: XZHENG@GAMMA.IS.TCU.EDU Subject: need help on peptide synthesizer To: methods_and_reagents@net.bio.net Message-id: <01I0S48DGZZ600DZMP@GAMMA.IS.TCU.EDU> X-VMS-To: IN%"methods_and_reagents@net.bio.net" MIME-version: 1.0 Content-type: TEXT/PLAIN; CHARSET=US-ASCII Content-transfer-encoding: 7BIT From: ISVMS3::XZHENG 3-FEB-1996 15:00:16.05 To: IN%"METHODS_AND_REAGENTS@NET.BIO.NET" CC: XZHENG Subj: NEED HELP ON PEPTIDE SYNTHESIZER Hi, netters: we are interested in getting an automatic peptide synthesizer. could you tell me the companies who sell the equipment and how much it will cost. any advise will be greatly appreciated. Alan Zheng Texas Christian Univ. Chemistry Dept. tel:(817) 921-7195 fax:(817) 921-7110 e-mail: XZHENG@gamma.is.tcu.edu --Boundary (ID 01MLeX23CE1DmndbOqTJAA)-- From owner-proteins@net.bio.net Fri Feb 02 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!vixen.cso.uiuc.edu!uwm.edu!psuvax1!news.eecs.nwu.edu!newsfeed.acns.nwu.edu!news.acns.nwu.edu!news From: alanhowe@merle.acns.nwu.edu (Alan K. Howe) Newsgroups: bionet.molbio.proteins Subject: Specific MAPK-K inhibitor Date: Fri, 02 Feb 96 19:14:03 EST Organization: Northwestern University, Chicago, IL. USA Lines: 18 Message-ID: <4eucas$itn@news.acns.nwu.edu> Reply-To: alanhowe@merle.acns.nwu.edu NNTP-Posting-Host: pc22.microbio.nwu.edu Mime-Version: 1.0 X-Newsreader: WinVN 0.93.9 There's an organic compound that was discovered by a group at Parke-Davis that specifically inhibits MEK (MAP kinase kinase) activity. It was described in PNAS, sometime in the middle or end of '95. My questions are two fold: 1.) Has anybody used this compound, and if so, what do you think? 2.) How did you get it? Can you order some from Parke-Davis, or did you have to write to the authors of the PNAS paper and wait a year for a response? Any practical information on this compound would be enormously appreciated. -- Alan K. Howe Northwestern University, Chicago, IL. USA alanhowe@merle.acns.nwu.edu From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!CS.SANDIA.GOV!wehart From: wehart@CS.SANDIA.GOV (William E. Hart) Newsgroups: bionet.molbio.proteins Subject: Update for Second SNL Workshop on Computational Molecular Biology Date: 4 Feb 1996 22:26:11 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 52 Sender: daemon@net.bio.net Distribution: world Message-ID: <9602050623.AA00633@kiva.cs.sandia.gov.noname> NNTP-Posting-Host: net.bio.net UPDATE*UPDATE**UPDATE*UPDATE*UPDATE**UPDATE*UPDATE*UPDATE $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ The Second Sandia National Laboratories Workshop on Computational Molecular Biology March 4-6, 1996 Albuquerque, New Mexico Organized in collaboration with DIMACS Special Year on Mathematical Support for Molecular Biology Funded by DOE MICS Office of Scientific Computing Applied Mathematics Program Dr. Fred Howes, Director Two issues of interest for potential participants: (1) A Poster Session was added to the program due to repeated suggestions from the participants. It will be scheduled on Monday March 4, 1996 Interested participants should send a ONE PAGE ABSTRACT via email to Susan Flores (sgflore@cs.sandia.gov) by February 17. A book of abstracts will be available at the workshop. (2) Information about the workshop could be obtained from the workshop web page http://www.cs.sandia.gov/cmb_workshop96.html (a previous announcement included a missprint in the web address) or contacting: Sorin Istrail, Workshop Chair Sandia National Laboratories Massively Parallel Computing Research Laboratory Algorithms and Discrete Mathematics Albuquerque, NM 87185-1110 Phone : (505) 845-7612 Secretary: (505) 845-7432 Fax : (505) 845-7442 Email : scistra@cs.sandia.gov www : http://www.cs.sandia.gov/~scistra/ From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.tamu.edu!news.utdallas.edu!news.swmed.edu!news From: "R. Bryan Sutton" Newsgroups: bionet.molbio.proteins Subject: Re: Beta-lactamase assay INSOLUBLE? Date: 5 Feb 1996 04:54:37 GMT Organization: University of Texas Southwestern Medical Center Lines: 10 Message-ID: <4f42id$1c7@swsu65.swmed.edu> References: <4e6ivq$36f@swsu65.swmed.edu> <51ka22tqyt.fsf_-_@shiva1-7.bms.com> NNTP-Posting-Host: dan.swmed.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; 68K) To: nathan@protos.bms.com X-URL: news:51ka22tqyt.fsf_-_@shiva1-7.bms.com We've used a compound called PADAC to test for beta-lactamase activity. It turns from a deep purple to clear when cleaved by bla. Calbiochem sells it. Bryan Sutton The University of Texas Southwestern Medical Center Dallas,TX email: sutton@howie.swmed.edu From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsserver.jvnc.net!synapse.bms.com!news-admin From: nathan@shiva1-7.bms.com (Nathan Siemers) Newsgroups: bionet.molbio.proteins Subject: Re: Beta-lactamase assay INSOLUBLE? Date: 04 Feb 1996 19:02:18 -0800 Organization: Bristol-Myers Squibb Lines: 10 Message-ID: <51ka22tqyt.fsf_-_@shiva1-7.bms.com> References: <4e6ivq$36f@swsu65.swmed.edu> Reply-To: nathan@protos.bms.com NNTP-Posting-Host: shiva1-7.bms.com X-Newsreader: September Gnus v0.26/Emacs 19.29 On a related note, does anyone know of a lactamase substrate that produces an Insoluble colored product? Something suitable for plaque lifts, histology, etc? I tried one method (no ref handy) using a cephalosporin with a thiol 3' substituent and added NBT (I think), but could not get it to work. nathan From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.tamu.edu!news.utdallas.edu!news.swmed.edu!news From: "R. Bryan Sutton" Newsgroups: bionet.molbio.proteins Subject: Re: Beta-lactamase assay INSOLUBLE? Date: 5 Feb 1996 05:04:09 GMT Organization: University of Texas Southwestern Medical Center Lines: 9 Message-ID: <4f4349$1c7@swsu65.swmed.edu> References: <4e6ivq$36f@swsu65.swmed.edu> <51ka22tqyt.fsf_-_@shiva1-7.bms.com> <4f42id$1c7@swsu65.swmed.edu> NNTP-Posting-Host: dan.swmed.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; 68K) X-URL: news:4f42id$1c7@swsu65.swmed.edu opps! I just realized that you wanted an INSOLUBLE product. soluble... insoluble.... Well, I was close.. Bryan Sutton The University of Texas Southwestern Medical Center Dallas,TX email: sutton@howie.swmed.edu From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!gatech!newsfeed.pitt.edu!bb3.andrew.cmu.edu!casaba.srv.cs.cmu.edu!das-news2.harvard.edu!fas-news.harvard.edu!fas.harvard.edu!rickles From: rickles@fas.harvard.edu (Richard Rickles) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: FENTOMOLE PROTEIN SEQUENCING Date: 4 Feb 1996 18:04:53 GMT Organization: Harvard University, Cambridge, Massachusetts Lines: 10 Message-ID: <4f2sg5$n62@decaxp.harvard.edu> NNTP-Posting-Host: fas.harvard.edu X-Newsreader: NN version 6.5.0 #3 (NOV) Xref: biosci bionet.molbio.methds-reagnts:39778 bionet.molbio.proteins:6927 FEMTOMOLE PROTEIN SEQUENCING-Means you CAN identify those proteins on PVDF from 2D gels and minigels... contact brauer@ariad.com (include your fax number) From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!tank.news.pipex.net!pipex!oleane!jussieu.fr!univ-lyon1.fr!in2p3.fr!swidir.switch.ch!swsbe6.switch.ch!scsing.switch.ch!news.belwue.de!fu-berlin.de!cs.tu-berlin.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins Subject: Re: Tissue protein isolation? Date: 4 Feb 1996 14:15:36 GMT Organization: University of Wuerzburg, Germany Lines: 22 Message-ID: <4f2f28$elm@winx03.informatik.uni-wuerzburg.de> References: NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Jeffrey Upperman (upperman@njmsa.UMDNJ.EDU) wrote: > does anyone have a protocol or reference for tissue protein isolation? > be gentle i am new to the protein world... It would be helpful if you could share with us what kind of protein you want to isolate, how you want to follow it during the course of isolation (i.e. can you detect it by activity, western blot, ...), whether you can follow an established protocol or not. For general reading, I would recommend Robert K. Scopes, "Protein Purification", 3rd edition ? , Springer. (I liked the 2nd ed. more, however.) Good luck! --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!usenet.eel.ufl.edu!warwick!bsmail!mail!cr5739 From: cr5739@mail.bris.ac.uk (CD Rees) Subject: Gamma glutaminyl-lysine? Message-ID: Sender: usenet@uns.bris.ac.uk (Usenet news owner) Nntp-Posting-Host: mail.bris.ac.uk Reply-To: cr5739@bristol.ac.uk Organization: University of Bristol, England X-Newsreader: TIN [version 1.2 PL1] Date: Sun, 4 Feb 1996 12:59:12 GMT Lines: 12 Could anybody please help me find any information on the amino acid gamma glutaminyl-lysine and the consequences of its deficiency? I have to do a presentation but I can't find any references in any of the standard textbooks. Any help would be gratefully received. Thanks. Chris Rees. From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!agate!howland.reston.ans.net!newsfeed.internetmci.com!news.compuserve.com!news.production.compuserve.com!news From: Pier Carlo Montecucchi <100143.765@CompuServe.COM> Newsgroups: bionet.biology.cardiovascular,bionet.molbio.proteins,sci.bio.technology,sci.med Subject: r-hirudin Date: 4 Feb 1996 09:39:00 GMT Organization: HI-TECH INDUSTRY CONSULTANTS Lines: 4 Distribution: inet Message-ID: <4f1urk$b2l$2@mhafc.production.compuserve.com> Xref: biosci bionet.biology.cardiovascular:772 bionet.molbio.proteins:6924 sci.bio.technology:4711 sci.med:109072 Searching a financial partner for the clinical development of the product Please send E mail to: 100143,765@compuserve.com Pier Carlo Montecucchi FL USA From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!agate!howland.reston.ans.net!nntp.coast.net!chi-news.cic.net!newsfeed.internetmci.com!news.compuserve.com!news.production.compuserve.com!news From: Pier Carlo Montecucchi <100143.765@CompuServe.COM> Newsgroups: bionet.biology.cardiovascular,bionet.molbio.proteins,sci.bio.technology,sci.med Subject: r-hirudin Date: 4 Feb 1996 09:36:24 GMT Organization: HI-TECH INDUSTRY CONSULTANTS Lines: 7 Distribution: inet Message-ID: <4f1umo$b2l$1@mhafc.production.compuserve.com> Xref: biosci bionet.biology.cardiovascular:771 bionet.molbio.proteins:6923 sci.bio.technology:4710 sci.med:109071 Searcing a financial partner for the clinical development of the product(r-hirudin). The pre-clinical, Phase I, Phase II studies are ready. All the dossiers are in compliance with EEC and FDA regulations and guidelines. Please send E mail to : 100143,765Compuserve.com P.C. Montecucchi From owner-proteins@net.bio.net Sat Feb 03 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!chi-news.cic.net!io.org!usenet From: khaddad@io.org (keith haddad) Newsgroups: bionet.molbio.proteins Subject: Pectin Date: Mon, 05 Feb 1996 02:43:01 GMT Organization: Internex Online, Toronto, Ontario, Canada (416 363 3783) Lines: 14 Message-ID: <4f3g3m$78n@ionews.io.org> NNTP-Posting-Host: dyna-60.net7a.io.org X-Newsreader: Forte Free Agent 1.0.82 I need to know how to extract pectinase from pectinolytic organism. What are the uses in industry for this enzyme in industry?? Well I have a good idea of what Im doing but Id like a second option. thanx Keith khaddad@io.org http://www.io.org/~khaddad ;) From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!daresbury!yama.mcc.ac.uk!news.york.ac.uk!news From: Graham Timmins Newsgroups: bionet.molbio.proteins Subject: Proteins in Tears? Date: 5 Feb 1996 13:52:31 GMT Organization: The University of York, UK Lines: 5 Message-ID: <4f522v$qgn@netty.york.ac.uk> NNTP-Posting-Host: chempc28.york.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.2N (Windows; I; 16bit) I would be grateful if anyone either knows or could point me in the direction of information on tear proteins, specifically a breakdown of what proteins and how much are present in tears (preferably human data). Thanks. From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!daresbury!yama.mcc.ac.uk!news.mcb.net!news.uni-stuttgart.de!news.rhrz.uni-bonn.de!RRZ.Uni-Koeln.DE!news.dfn.de!uni-muenster.de!news From: schille@uni-muenster.de Newsgroups: bionet.molbio.proteins Subject: non radioactiv cAMP measurement Date: 5 Feb 1996 13:15:42 GMT Organization: Westfaelische Wilhelms-Universitaet Muenster, Germany Lines: 6 Message-ID: <4f4vtu$ido@majestix.uni-muenster.de> NNTP-Posting-Host: fplc.uni-muenster.de X-Newsreader: AIR News 3.X (SPRY, Inc.) How could i measure cAMP with a non radioactiv method? Does anybody know about intracellular response of HUVEC (human umbilical vein endothelial cells) after stimulation with the coagulations enzyme factor IX or factor IXa ? Hermann Schillers From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!GAMMA.IS.TCU.EDU!XZHENG From: XZHENG@GAMMA.IS.TCU.EDU Newsgroups: bionet.molbio.proteins Subject: $10,000-$20,000 for a peptide synthesizer? Date: 5 Feb 1996 14:55:42 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I0V0K5Z8FM00DYSE@GAMMA.IS.TCU.EDU> NNTP-Posting-Host: net.bio.net Hi! Is there a way to get a automatic peptide synthesizer below $20,000? any suggestion about the company who sell it will be greatly appreciated! Alan Zheng Chemistry Dept. Texas Christian Univ. tel: (817) 921-7195 fax: (817) 921-7110 From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!nntp.coast.net!news00.sunet.se!sunic!news99.sunet.se!news.uni-c.dk!news From: vadtsvet@biochem.ou.dk (Vadim Tsvetnitsky) Newsgroups: bionet.molbio.proteins Subject: Protein analysis software Date: 5 Feb 1996 09:18:30 GMT Organization: Odense University, DENMARK Lines: 12 Message-ID: <4f4i16$4rl@news.uni-c.dk> NNTP-Posting-Host: vadim.biochem.ou.dk X-Newsreader: WinVN 0.92.6+ There is an excellent program written by Peter Hojrup, Odense University, Denmark called General Protein/Mass analysis for Windows (GPMAW). It is very useful for the most of protein analysis including generation of proteolytic peptides, pI, mass-spec, hydrophobicity plots, secondary structure and so on. Can read EMBL/Atlas data from CD-ROM or accept yout sequence. For more info and licensing terms contact: Peter Højrup e-mail: php@pr-group.ou.dk I use this software and can really recommend it. From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!newsfeed.internetmci.com!info.ucla.edu!nnrp.info.ucla.edu!usenet From: Duilio Cascio Newsgroups: bionet.molbio.proteins Subject: NEW UCLA-FACULTY POSITION Date: Mon, 05 Feb 1996 17:54:18 -0800 Organization: University of California, Los Angeles Lines: 30 Message-ID: <3116B4CA.167E@ucla.edu> NNTP-Posting-Host: galileo.mbi.ucla.edu Mime-Version: 1.0 Content-Type: text/html; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b6a (X11; I; OSF1 V3.2 alpha) Content-Disposition: inline; filename="doeposition.html" UCLA FACULTY POSITION UCLA-DOE LABORATORY OF STRUCTURAL BIOLOGY AND MOLECULAR MEDICINE The division of Structural Biology and Genetics of the UCLA-DOE Laboratory of Structural Biology and Molecular Medicine will appoint one faculty position at the level of Assistant Professor or Assistant Professor in Residence, jointly with departments in the College of Letters and Science or the School of Medicine. We seek a computational biochemist with an interest in genome analysis. Truly exceptional candidates in related areas will also be considered. Set up and research funds will come from the UCLA-DOE Lab. Candidates must have significant research accomplishments, the ability to establish a vigorous research program, and a commitment to excellence in education. UCLA is an Equal Opportunity Employer and encourages applications from women and minority candidates. Applicants should provide a curriculum vitae, a summary of proposed research, copies of key publications or preprints, and three letters of recommendation by March 15, 1996 (extended deadline). Applications should be sent to: Dr. David Eisenberg Chair of Search Committee UCLA-DOE Lab UCLA, Box 951570 Los Angeles, CA 90095-1570 FAX: (310) 206-3914. From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!swrinde!cssun.mathcs.emory.edu!news.cc.emory.edu!usenet From: skris01@unix.cc.emory.edu (Krishnaswamy) Newsgroups: bionet.molbio.proteins Subject: Re: APMSF Date: Mon, 05 Feb 1996 23:23:06 GMT Organization: Emory University Lines: 26 Message-ID: <4f63i0$cb9@moe.cc.emory.edu> References: <4dmhgp$ndd@newsbf02.news.aol.com> NNTP-Posting-Host: skrish.hematology.emory.edu X-Newsreader: Forte Free Agent 1.0.82 qualichem@aol.com (QUALICHEM) wrote: >ANYBODY HAVE ANY EXPERIENCE/SUCCESS WITH A PROTEASE INHIBITOR CALLED >APMSF? WHERE DO YOU GET IT, HOW MUCH DO YOU USE, ETC. THANKS APMSF=amidino phenylmethanesulphonyl fluoride This is a serine proteinase inhibitor with added specificity for Arg specific serine proteinases afforded by the amidino moiety. For example, APMSF does not inhibit chymotrypsin significantly. The original citation for this compound is: Laura, R., Robison,D.J. and Bing, D.H. p-Amidinophenyl methanesulphonyl fluoride, an irrversible inhibitor of serine proteases. (1980) Biochemistry 19, 4859-4864 Many of us working with coagulation proteases use APMSF as it reacts rapidly and irreversibly with arginine-specific serine proteinases such as thrombin and factor Xa and is also very unstable at pH>=7. Thus, the problems with excess inhibitor removal are greatly minimized. The amounts you will need to use will depend on the protease you are trying to inhibit, the protease concentration and the pH at which you want to work at. From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!torn!utnut!oci!news From: Flip Hoedemaeker Subject: Re: Proteins in Tears? Content-Type: text/plain; charset=us-ascii Message-ID: <311687DF.41C6@oci.utoronto.ca> Sender: news@oci.utoronto.ca Content-Transfer-Encoding: 7bit Organization: Ontario Cancer Institute - U of Toronto References: <4f522v$qgn@netty.york.ac.uk> Mime-Version: 1.0 Date: Mon, 5 Feb 1996 22:42:39 GMT X-Mailer: Mozilla 2.0b6a (X11; I; IRIX 5.2 IP20) Lines: 1 Just out of my head, tears contain lot's of lysozyme.... From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!GAMMA.IS.TCU.EDU!XZHENG From: XZHENG@GAMMA.IS.TCU.EDU Newsgroups: bionet.molbio.proteins Subject: $10,000-$20,000 for a peptide synthesizer? Date: 5 Feb 1996 14:57:19 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I0V0R2LCC200DYSE@GAMMA.IS.TCU.EDU> NNTP-Posting-Host: net.bio.net From: ISVMS3::XZHENG 5-FEB-1996 16:53:24.81 To: IN%"proteins@net.bio.net" CC: XZHENG Subj: $10,000-$20,000 for a peptide synthesizer? Hi! Is there a way to get a automatic peptide synthesizer below $20,000? any suggestion about the company who sell it will be greatly appreciated! Alan Zheng Chemistry Dept. Texas Christian Univ. tel: (817) 921-7195 fax: (817) 921-7110 From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!tank.news.pipex.net!pipex!lade.news.pipex.net!pipex!tube.news.pipex.net!pipex!dish.news.pipex.net!pipex!news00.sunet.se!sunic!news99.sunet.se!news.uni-c.dk!news From: Oliver Politz Newsgroups: bionet.molbio.proteins Subject: Re: removing nucleic acids from extracts Date: Mon, 05 Feb 1996 18:37:27 -0800 Organization: Carlsberg Laboratory, Copenhagen, DK Lines: 23 Message-ID: <3116BEE6.20F1@biobase.dk> References: <310CD910.4A12@quickmail.ucsf.edu> <4ekma4$bln@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: lgbppp33.uni-c.dk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b6a (Win16; I) Cornelius Krasel wrote: > > trey simmons (trey_simmons.gicd@quickmail.ucsf.edu) wrote: > > I know there must be a straightforward way to remove nucleic acids > > from extracts, but I'm having trouble finding one. > > I think you can precipitate nucleic acids specifically with streptomycin > sulphate. A reference escapes me at the moment. > > Good luck! > > --Cornelius. > > -- > /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ > /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ > /* "Science is the game we play with God to find out what His rules are." */ I f you want get away any nucleic acid try BENZONASE supplied by Merck ( I am not involved in this company) it is really a god enzyme wich degrades RNA and DNA quit fast. We usually use it for protein preparations. Good luck Oliver Politz From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!GAMMA.IS.TCU.EDU!XZHENG From: XZHENG@GAMMA.IS.TCU.EDU Newsgroups: bionet.molbio.proteins Subject: address of ABI (Perkin Elmer)? Date: 5 Feb 1996 14:49:01 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 9 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I0V0H8CYSI00DYSE@GAMMA.IS.TCU.EDU> NNTP-Posting-Host: net.bio.net From: ISVMS3::XZHENG 5-FEB-1996 16:45:27.06 To: IN%"proteins@net.bio.net" CC: XZHENG Subj: address of ABI (Perkin Elmer)? Hi, netters! Can anyone give me the address or telephone number of ABI (Perkin Elmer)? we areinterested in their products. thank you in advance! Alan Zheng From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!GAMMA.IS.TCU.EDU!XZHENG From: XZHENG@GAMMA.IS.TCU.EDU Newsgroups: bionet.molbio.proteins Subject: address of ABI (Perkin Elmer)? Date: 5 Feb 1996 14:47:45 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 4 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I0V0BOK8SY00DYSE@GAMMA.IS.TCU.EDU> NNTP-Posting-Host: net.bio.net Hi, netters! Can anyone give me the address or telephone number of ABI (Perkin Elmer)? we areinterested in their products. thank you in advance! Alan Zheng From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!gatech!concert!bigblue.oit.unc.edu!mmlds1.pha.unc.edu!iiv From: Iosif Vaisman Newsgroups: bionet.biophysics,bionet.molec-model,bionet.xtallography,bionet.molbio.proteins,bionet.structural-nmr Subject: Gordon Conference on Water and Aqueous Solutions Date: Mon, 5 Feb 1996 11:36:42 -0500 Organization: The University of North Carolina at Chapel Hill Lines: 158 Message-ID: NNTP-Posting-Host: mmlds1.pha.unc.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Xref: biosci bionet.biophysics:1679 bionet.molec-model:804 bionet.xtallography:2352 bionet.molbio.proteins:6937 bionet.structural-nmr:1088 Crossposted from Water Science Network (water@listserv.unc.edu) Gordon Research Conference PHYSICS AND PHYSICAL CHEMISTRY OF WATER AND AQUEOUS SOLUTIONS Aug 4 - Aug 9, 1996 ------------------------------------------------------ ------------------------------------------------------ George E. Walrafen, Chair H. Eugene Stanley, Vice Chair ------------------------------------------------------ ------------------------------------------------------ WATER IN PROTEINS--------------(Sunday evening, 4 Aug) A. Parsegian-------------------(Session chair) P. Nicholls/P. Kahn/R. Wolfenden ------------------------------------------------------ AMORPHOUS ICE TO HOT WATER----(Monday morning, 5 Aug) J. Teixeira--------------------(Session chair) G. Johari/O. Mishima/ A. Geiger/M. Neumann/ P. Debenedetti-----------------(Commentator) SCATTERING: X-RAY, NEUTRON-----(Monday evening, 5 Aug) M.-C. Bellissent-Funel---------(Session chair) T. Yamaguchi/A. Soper PLENARY POSTER SESSION #1 H. E. Stanley------------------(Poster chair) ------------------------------------------------------ ULTRAFAST DYNAMICS/HOLE BURNING-(Tuesday morning, 6 Aug) A. Laubereau-------------------(Session chair) E. Castner/R. J. Dwayne-Miller/ G. J. Small/H. Graener/ W. T. Lotshaw------------------(Commentator) LOW-FREQUENCY RAMAN SPECTROSCOPY--(Tuesday evening, 6 Aug) Y. Tominaga--------------------(Session chair) Y. C. Chu/Y. Kameda/ O. Faurskov-Nielsen------------(Commentator) ------------------------------------------------------- HYDRATION OF BIOMOLECULES------(Wednesday morning, 7 Aug) B. Schoenborn------------------(Session chair) S. Leikin/H. Berman/ A. Gronenborn/E. Mayer ICES AND CAGES-----------------(Wednesday evening, 7 Aug) P. Kusalik---------------------(Session chair) I. Svishchev/S.-H. Chen PLEANARY POSTER SESSION #2 H. E. Stanley------------------(Poster chair) ------------------------------------------------------- STRUCTURE AND DYNAMICS OF H-BONDED SYSTEMS---------------(Thursday morning, 8 Aug) C. A. Angell-------------------(Session chair) A. Luzar/T. Head-Gordon/ R. Saykally/N. Agmon THURSDAY EVENING: BANQUET, BUSINESS SESSION, ELECTION OF VICE CHAIR, ETC. CRITICAL PHENOMENA-------------(Thursday evening, 8 Aug) G. E. Walrafen-----------------(Session chair) B. Widom-----------------------(45 minute special speaker) Professor Widom's lecture will be a wide overview of the developments in CRITICAL PHENOMENA from the early days of the subject to the most recent exciting developments, such as renormalization group theory, etc. ----------------END OF CONFERENCE------------------------ THE POSTERS WILL BE ON DISPLAY ALL WEEK LONG!!! INFORMATION: Tentative poster titles for Gene Stanley's PLENARY POSTER SESSIONS, should be sent by E-mail or FAX directly to Gene (HES@BU.EDU, 617-353-3783). These poster sessions will be held on Monday and Wednesday evenings. The posters will be on display all week long, and they will be displayed in the big, well-lighted, red school house. The poster boards are 4 ft. X 8 ft., so there will be plenty of room for your material. More than one poster per person is also possible. We also want to emphasize as strongly as we can that EARLY REGISTRATION is absolutely essential. No applicants will be accepted by the GRC after about 135 people have registered!!!! Contact the GRC office; phone (401)-783-4011 or -3372; FAX (401)-783-7644; E-MAIL grc@grcmail.grc.uri.edu to obtain your registration forms. The GRC has streamlined the registration procedures. For example, they can take registration payments by FAX if you give your credit card number on their form, sign it, and FAX it back. We will also mail out some registration forms to people who are definitely attending, or to people who attended in 1994. However, it is best to get your materials yourself from the GRC people. We plan mini- or ground-zero review for most sessions. These will be presented by the session chairs. The ground-zero reviews will involve overviews of the talks to follow in each session. The ground- zero reviews WILL NOT involve talks by the session chairs about their own work. The ground-zero reviews are intended to introduce each session for people in the audience who may not be familiar with the previous work in the area of the session. We will also have two micro-reviews Monday evening and Wednesday evening to save time for the poster sessions to follow. In addition we will have commentators (about 5 minutes) following the sessions on Monday morning, Tuesday morning, and Wednesday evening. The purpose of the commentators is to present a summing up, as well as to indicate problems, or regions of work which should be exploited in the future. Our Thusday evening speaker, after the banquet, will be Professor Ben Widom from Cornell University. This will really be a special treat for all of us. Professor Widom will tell us about developments in "CRITICAL PHENOMENA" from the past to the present. Our mixer will be held Sunday afternoon starting roughly at 4:30 or 5:00 p.m. before the full dinner on Sunday evening. This year you will get a dinner, and not a mini-dinner, on Sunday evening. Gene Stanley and I hope to greet all of you in person at the mixer. Adrian Parsegian is helping us with the reception, and he is also the first session chair, Sunday night. We could hardly have gotten a better person for both assignments. He has assured me that he will pick up the liquid refreshments for us on his way to the conference. Remember, REGISTER SOON. We hope to see you in August. Sincerely yours, George Walrafen Chair Ph:(202)-806-6897 Fax:(202)-806-5475 e-mail:YCC@SCS.HOWARD.EDU H. Eugene Stanley Vice Chair Ph:(617)-353-2617 FAX:(617)-353-3783 e-mail:HES@BU.EDU From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!fdn.fr!jussieu.fr!univ-poitiers.fr!usenet From: volz Newsgroups: bionet.molbio.proteins Subject: Computation of proteins Date: 5 Feb 1996 16:30:12 GMT Organization: C.R.I.U.P., Universite de Poitiers (FRANCE) Lines: 3 Message-ID: <4f5bak$gpa@hermes.univ-poitiers.fr> NNTP-Posting-Host: pcseb.univ-poitiers.fr Who could give me informations about computer simulation of proteins? (Works, software. labs. Searcher...). THank you From owner-proteins@net.bio.net Sun Feb 04 22:00:00 1996 Path: biosci!FREENET.TORONTO.ON.CA!bz182 From: bz182@FREENET.TORONTO.ON.CA (Wei Hu) Newsgroups: bionet.molbio.proteins Subject: gfp sensitivity Date: 5 Feb 1996 20:12:33 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net hi there, i don't know whether gfp can be used to trace a low abundance protein in mammalian cells, which is usually pulse-labeled with 35s-met. could anyone experienced in gfp please tell me the related information. many thanks in advance. wei From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!vixen.cso.uiuc.edu!usenet.ucs.indiana.edu!bronze.ucs.indiana.edu!lruble From: lruble@bronze.ucs.indiana.edu (the End) Newsgroups: bionet.molbio.proteins,bionet.microbiology Subject: GroEL as RNA chaperone? Date: 7 Feb 96 06:15:59 GMT Organization: Indiana University, Bloomington Lines: 16 Message-ID: NNTP-Posting-Host: bronze.ucs.indiana.edu NNTP-Posting-User: lruble X-Newsreader: NN version 6.5.0 #5 (NOV) Xref: biosci bionet.molbio.proteins:6961 bionet.microbiology:4867 How well is the proposal of Georgellis that GroEL functions as a chaperone for mRNA going over with everyone? I have some catching up to do, but am first disturbed by the lack of evidence for interaction with RNAs other than OmpA and 9S in Molecular Micro 16, p.1259 (1995). I like the study, but feel like what has been shown is very preliminary, and difficult to draw such a model from. Anyone? (Please E-mail me a copy of responses) Jim J. Graham Biology Department Washington University of St. Louis From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!biochem.kth.se!mostafa From: mostafa@biochem.kth.se (Mostafa Ronaghi) Newsgroups: bionet.molbio.proteins Subject: immobilization Date: 6 Feb 1996 02:03:51 -0800 Organization: Biochemistry, Royal Institute of Technology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: <3117269E.167EB0E7@biochem.kth.se> NNTP-Posting-Host: net.bio.net Dear all: Could anyone tell me which immobilization techniq gives the highest binding capacity in an area of cm2. The immobilization of streptavidin will be performed in a transparent glass tube or in a capillary. Thanks in advance Best Regards, mostafa@biochem.kth.se From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!primus.ac.net!news.cais.net!nntp.uio.no!nntp.uib.no!nntp-bergen.UNINETT.no!nntp-trd.UNINETT.no!due.unit.no!usenet From: Hans Stenoien Newsgroups: bionet.molbio.proteins Subject: PAL/anthocyanin-3-glucocyltransferase Date: Tue, 06 Feb 1996 09:25:05 +0100 Organization: The Norwegian University of Science and Technology Lines: 6 Message-ID: <31171061.22B3@vm.unit.no> NNTP-Posting-Host: vm237.ubt.unit.no Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b6a (Win95; I) CC: hans.stenoien@vm.unit.no Does anyone know if there has been done any electrophoresis on PAL and/or anthocyanin-3-glucocyltransferase? I am interested in staining techniques for these enzymes. Thanks in advance! Hans Stenoien From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.uoregon.edu!cyberbro.uoregon.edu!user From: meneghini@molbio.uoregon.edu (silverback) Newsgroups: bionet.molbio.proteins,bionet.molbio.yeast,bionet.molbio.molec-model Subject: arrested nascent polypeptides Date: Tue, 06 Feb 1996 16:24:39 -0800 Organization: University of Oregon Lines: 12 Message-ID: NNTP-Posting-Host: cyberbro.uoregon.edu Xref: biosci bionet.molbio.proteins:6959 bionet.molbio.yeast:4692 o.k. does anyone know at which site (A or P) in the ribosome arrested nascent chains lacking stop codons accumulate? Logic tells me that in the absence of further message, the peptidyl tRNA may stay in the A site and not translocate to the P site. However puromycin is capable of releasing these nascent chains and the lore is (as I undersatnd it) that puromycin only releases nascent chains in the P site. Please help, this information is critical to my comprehensive exam proposals and no one in the literature has offered any information on this. thanks, marc meneghini Institute of molecular biology, university of oregon From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!lll-winken.llnl.gov!fnnews.fnal.gov!unixhub!news.Stanford.EDU!usenet From: Ignacio Marin Newsgroups: bionet.molbio.proteins Subject: HISTONE ACETYLASES/DEACETYLASES Date: 6 Feb 1996 22:22:20 GMT Organization: Stanford University Lines: 7 Message-ID: <4f8kas$sma@nntp.Stanford.EDU> NNTP-Posting-Host: bio-bb2.stanford.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; 68K) X-URL: news:bionet.molbio.proteins Is there any information available about clones histone acetylases/deacetylases in any organism?. Any information will be welcome. Ignacio Marin. Dpt. Biological Sciences. Stanford University. From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!howland.reston.ans.net!nntp.coast.net!zombie.ncsc.mil!news.missouri.edu!NewsWatcher!user From: 71242.3650@compuserve.com (Timothy J. Strabala) Newsgroups: bionet.molbio.proteins Subject: Re: serial immunoprecipitation Date: Tue, 06 Feb 1996 16:14:59 -0600 Organization: University of Missouri-Columbia Lines: 47 Distribution: world Message-ID: <71242.3650-0602961614590001@128.206.90.48> References: <9602022024.AA02236@mani.cbs.umn.edu> NNTP-Posting-Host: 128.206.90.48 In article <9602022024.AA02236@mani.cbs.umn.edu>, nora@biosci.cbs.umn.edu wrote: > Does anyone have experience with co-immunoprecipitating proteins, > solubilizing the proteins, and reacting them with another antibody? > We have tried solubilizing an immunoprecipitate with 4% SDS, diluting > it to 0.1% SDS for a subsequent antibody reaction, but this did not > produce positive results. We will now try a final concentration of > 0.05% SDS, as well as different concentrations of urea (using either > 8 M or 4 M for the initial solubilization). > > > If you have any experience with this approach or other reagents that > have worked, I would really appreciate learning of them. Thanks very > much. > > Nora Plesofsky-Vig > nora@biosci.cbs.umn.edu Hi Nora, It seems to me that your problems are likely due to high SDS concentrations denaturing your second antibody. What you might try is solubilizing your immunoprecipitated protein in 1% SDS, followed by dilution of the solubilized protein with 6 volumes of 50 mM Tris-HCl, pH 7.5 (25 C) 150 mM NaCl and 1% (v/v) Triton X-100 to sequester free SDS. Chill 10 min, spin 48000xg for 10 min. You should be able to immunoprecipitate the protein of interest out of the supernatant with your 2nd antibody, provided it recognizes the protein in a denatured state. Needless to say, since you are diluting a fair amount, you want to solubilize initially in the minimum volume you can get away with. I have also found that overnight immunoprecipitation using a Labquake shaker at 4 degrees C was very helpful for my purposes. This protocol is taken from from Vierstra & Quail (1982) PNAS 79 5272-5276. I have used it to extract and immunoprecipitate low abundance proteins from soybean hypocotyl sections, but I see no reason as to why this particular step couldn't be adapted to help solve your problem as well. I hope you find this helpful. Please feel free to e-mail me if you have any other questions. Good luck, Tim -- Tim Strabala University of Missouri-Columbia Dept. of Biochemistry 117 Schweitzer Hall Columbia, MO 65211 71242.3650@compuserve.com From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!howland.reston.ans.net!nntp.coast.net!col.hp.com!news.cis.okstate.edu!usenet From: Ron Tate Newsgroups: bionet.molbio.proteins Subject: Re: Tissue protein isolation? Date: Tue, 06 Feb 1996 14:51:55 -0600 Organization: Dept. of Biochemistry and Molecular Biology, OSU Lines: 28 Message-ID: <3117BF6B.2725@bmb-fs1.biochem.okstate.edu> References: NNTP-Posting-Host: firefly3.biochem.okstate.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b5 (Win95; I) Jeffrey Upperman wrote: > > does anyone have a protocol or reference for tissue protein isolation? > be gentle i am new to the protein world... > -- > Jeffrey S. Upperman,MD Internet: upperman@umdnj.edu > Research Fellow Voice: (201) 982-4474 > Dept. Anatomy, Cell Biology & Injury Fax: (201) 982-6803 > Dept. of Surgery Beeper: (201) 708-4150 > New Jersey Medical School Office: (201) 982-5045 > 185 S. Orange Ave., G606 Lab: (201) 982-5404 > Newark, NJ 07103 Jeffrey, For starters check out "Protein Purification; Principals and Practice" by Robert Scopes published by Springer-Verlag and volume 182 of the "Methods in Enzymology" series. They both will give you guidelines and techniques from tissue or cell disruption through almost any protein purification technique you could want. Happy Hunting -- Ron Tate Lab of Franklin Leach Dept. of Biochem. & Molecular Biology Oklahoma State University From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!sgigate.sgi.com!uhog.mit.edu!news.mtholyoke.edu!news.umass.edu!nic.umass.edu!sbl1.micro.umass.edu!greguera From: greguera@microbio.umass.edu (Gemma) Newsgroups: bionet.molbio.proteins Subject: CarboxyMethyl Chitin Date: Tue, 6 Feb 1996 11:38:18 Organization: Dept of Microbio. Lines: 13 Message-ID: NNTP-Posting-Host: sbl1.micro.umass.edu Keywords: Chitinase Substrates X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Hi everybody! I once asked in this group for advices regarding chitinase assays. I remembered someone told me he was using Carboxy-Methyl-Chitin as chitinase substrate. I have been looking for the reference he gave me in my files but I cannot find it. I do remember that this commercial CM-chitin had been supplied by some German laboratory but that is all I remember. I would strongly appreciate if you can tell me where I could find this type of chitinase substrate. Feel free to give me any advice you might consider helpful. Thanks in advance! Gemma From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!swrinde!sgigate.sgi.com!uhog.mit.edu!news.mtholyoke.edu!news.umass.edu!nic.umass.edu!sbl1.micro.umass.edu!greguera From: greguera@microbio.umass.edu (Gemma) Newsgroups: bionet.molbio.proteins Subject: CMC-chitin Date: Tue, 6 Feb 1996 11:27:13 Organization: Dept of Microbio. Lines: 1 Message-ID: NNTP-Posting-Host: sbl1.micro.umass.edu Keywords: chitinase substrates X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!primus.ac.net!news.cais.net!nntp.uio.no!biotek19 From: damartin@bioslave.uio.no (David Martin) Newsgroups: bionet.molbio.proteins Subject: Re: Is there any limitation of using BioRad method to..? Date: Tue, 06 Feb 96 12:07:14 GMT Organization: Biotechnology Centre of Oslo Lines: 32 Message-ID: <4f7ga5$5o3@ratatosk.uio.no> References: <4f7bu5$9ls@hpg30a.csc.cuhk.hk> NNTP-Posting-Host: biotek19.uio.no X-Newsreader: News Xpress Version 1.0 Beta #3 In article <4f7bu5$9ls@hpg30a.csc.cuhk.hk>, s935537@acs.csc.cuhk.hk (Ricky Leung) wrote: -->Hi everyone, --> --> Yesterday, I used BioRad method ( Barford method ) for -->determination of protein concentration. But precipitation occured -->after adding the dye. How long after? and exactly how did you do the reaction? What is in your protein buffer? -->I wonder to know if there is limitation of -->using such method to determine protein concentration in high salt -->buffer. The reagent does precipitate slowly on standing, so mix and measure the OD as soon as possible. Check the 'compatible reagents' chart (p337 of the '96 catalogue) ..David ========================================================= * David Martin, PhD - Post-Doctoral Research Fellow * * Atherosclerosis and Thrombosis research group * * Biotechnologisenteret i Oslo * * Gaustadalleen 21 (Postboks 1125 Blindern) * * N-0316 Oslo * * Norway * * Tel: +47 22 95 84 54 Fax: +47 22 69 41 30 * * email: david.martin@biotek.uio.no * ========================================================= From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!gossip.pyramid.com!news.sedona.net!usenet From: Ann Rathbun Newsgroups: bionet.molbio.proteins Subject: Analytical Biochem Dev't Manager Date: 5 Feb 1996 19:13:20 GMT Organization: Sedona Internet Services, Inc. Lines: 52 Message-ID: <4f5ksg$6ku@labrat.sedona.net> NNTP-Posting-Host: client8.sedona.net Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Mailer: Mozilla 1.2N (Windows; I; 16bit) Manager, Analytical Biochemistry Development We are seeking a Manager for a newly formed Methods Development group with our biopharmaceutical client company on the East Coast. The successful Ph.D.-level Protein Biochemist will be responsible for methods development for the characterization and evaluation of [protein] products and processes for our biopharmaceutical client company. Candidates must have exceptionally strong communication skills as he/she will be the key link with the company’s foreign collaborators and will be responsible for putting forth ideas and plans to implement their various projects. Skills/Background: · Ph.D. in Biochemistry, Analytical Chemistry or related discipline; · Analytical methods development for proteins, esp. recombinant; · Strong Mass Spec experience with is a requirement; · 4 plus years postdoc with supervisory [hiring, promotion and firing responsibilities] experience; · Industry experience is preferred; · Ideally this person will have GMP and Regulatory filing experience. Other Desired Skills: Amino Acid Analysis Bioassay Chromatography Circular Dichroism Electrophoresis HPLC Sequencing, N- & C- terminal Structure/Function Tryptic Mapping If you have an interest in this or other opportunities, please mail or FAX your CV/resume to RS&A to the attention of Ann G. Rathbun, Managing Director. All correspondence is held in strict confidence. Ann Rathbun Managing Director Rathbun, Sapir & Associates P.O. Box 2337 Sedona, AZ 86339-2337 * USA (520) 284-3360 Office (520) 284-3361 FAX E-mail: rathbun@ sedona.net From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!primus.ac.net!news.cais.net!xara.net!peer-news.britain.eu.net!sunsite.doc.ic.ac.uk!hgmp.mrc.ac.uk!ebi.ac.uk!higgins From: higgins@ebi.ac.uk Subject: Re: WHO KNOWS THE RATE OF TRANSLATION??!! (AA/min) Sender: news@ebi.ac.uk (Mr news) Message-ID: <1996Feb5.164559@ebi.ac.uk> Date: Mon, 5 Feb 1996 16:45:59 GMT Lines: 20 References: <4e5hga$sis@rc1.vub.ac.be> <1996Jan31.122553@ebi.ac.uk> <4epv98$jim@winx03.informatik.uni-wuerzburg.de> Organization: European BioInformatics Institute In article <4epv98$jim@winx03.informatik.uni-wuerzburg.de>, krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) writes: > higgins@ebi.ac.uk wrote: >> (I have no idea if there is a Genes V) > > Yes, there is. (However, I liked IV better :-) > > --Cornelius. > So that was why it was so cheap :-). (I found it in a local charity bookshop (Oxfam)). Des P.S. there is a deliberate mistake in my last posting on this thread. > -- > /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ > /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ > /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Path: biosci!agate!hpg30a.csc.cuhk.hk!news.cuhk.edu.hk!s935537 From: s935537@acs.csc.cuhk.hk (Ricky Leung) Newsgroups: bionet.molbio.proteins Subject: Is there any limitation of using BioRad method to..? Date: 6 Feb 1996 10:52:53 GMT Organization: Chinese University of Hong Kong Lines: 13 Message-ID: <4f7bu5$9ls@hpg30a.csc.cuhk.hk> Reply-To: gene@cuhk.hk NNTP-Posting-Host: s935537@hp735b.csc.cuhk.hk X-Newsreader: TIN [version 1.2 PL2] Hi everyone, Yesterday, I used BioRad method ( Barford method ) for determination of protein concentration. But precipitation occured after adding the dye. I wonder to know if there is limitation of using such method to determine protein concentration in high salt buffer. What causes the problem? How can I solve it? Could anyone suggest methods to improve the situation? Thanks in advance. Ricky Leung From owner-proteins@net.bio.net Mon Feb 05 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!torn!nott!cunews!wabakimi!nbatada From: nbatada@chat.carleton.ca (Nizar Batada) Subject: BIOCHM STUDENT--> WORK IN CALIF AREA X-Nntp-Posting-Host: wabakimi.carleton.ca Message-ID: Sender: news@cunews.carleton.ca (News Administrator) Organization: Carleton University X-Newsreader: TIN [version 1.2 PL2] Date: Wed, 7 Feb 1996 02:15:31 GMT Lines: 20 Hi all, I am a third year undergraduate student in Honours Biochemistry looking for a position as a research assistant (for summer 96) in areas around San Jose, San francisco, or areas within an hours drive from them. I have a 3.78 G.P.A and am planning to pursue masters or PhD in Molecular genetics or Biochemistry. Although I would like to get paid for the work, I would consider working without pay. If anyone reading this knows or is able to offer such a position please drop me an email. A more detailed CV and reference is available on request. Thanks, ---------------------------------------------------------------------- Nizar Batada email address: nbatada@chat.carleton.ca ---------------------------------------------------------------------- From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!oleane!jussieu.fr!univ-lyon1.fr!in2p3.fr!swidir.switch.ch!swsbe6.switch.ch!scsing.switch.ch!news.belwue.de!fu-berlin.de!arnold.chemie.fu-berlin.DE!not-for-mail From: saihtam@chemie.fu-berlin.de (Mathias Dreger) Newsgroups: bionet.molbio.proteins Subject: Affinity purification of tyrosine phosphorylated proteins Date: Wed, 07 Feb 96 15:51:41 GMT Organization: FU Berlin Lines: 11 Message-ID: <4f9t1j$ihr@fu-berlin.de> NNTP-Posting-Host: arnold.chemie.fu-berlin.de (160.45.28.9) X-Access: 16 17 19 X-Newsreader: News Xpress Version 1.0 Beta #4 Hello there, At present, I try to purify tyrosine-phosphorylated proteins using protein A-sepharose coupled mAb PY 20 as affinity matrix. Unfortunately, yields are absolutely bad. So I ask for advice from people who have some experience with this topic especially concerning the amount of starting material needed and good procedures to elute the affinity column. It would also be nice if somebody had some useful references. Thanks in advance Mathias From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!agate!hpg30a.csc.cuhk.hk!news.cuhk.edu.hk!s935537 From: s935537@acs.csc.cuhk.hk (Ricky Leung) Newsgroups: bionet.molbio.proteins Subject: Re: Is there any limitation of using BioRad method to..? Date: 7 Feb 1996 13:37:10 GMT Organization: Chinese University of Hong Kong Lines: 10 Message-ID: <4fa9u6$o2d@hpg30a.csc.cuhk.hk> References: <4f7bu5$9ls@hpg30a.csc.cuhk.hk> Reply-To: gene@cuhk.hk NNTP-Posting-Host: s935537@hp735a.csc.cuhk.hk X-Newsreader: TIN [version 1.2 PL2] Hi everyone, All suggestions are welcome One additional information I have to write is the sample was dissolved in potassium chloride and sodium cholate. Was the precipitation related to sodium cholate? Ricky Leung From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!vixen.cso.uiuc.edu!newsrelay.iastate.edu!newsxfer.itd.umich.edu!nntp.cs.ubc.ca!cs.ubc.ca!unixg.ubc.ca!news From: yossi@unixg.ubc.ca Newsgroups: bionet.molbio.proteins Subject: Cholesterol Oxidase Date: Wed, 07 Feb 1996 08:22:55 GMT Organization: The University of British Columbia Lines: 7 Message-ID: <4f9mki$jla@nntp.ucs.ubc.ca> NNTP-Posting-Host: port04.annex4.net.ubc.ca X-Newsreader: Forte Free Agent 1.0.82 I am trying to mesure chod activity. However, Cholesterol due to its insouluble form in water gives high and unstable background. Meanwhile I am using the recomended assay at 240nm (the formation of 3-cholestane from cholesterol) using low triton x-100 and phosphate buffer. Any suggestions? thanks Yossi From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!newsfeed.internetmci.com!vixen.cso.uiuc.edu!news.ksu.ksu.edu!news.cis.okstate.edu!usenet From: Ann Williams Newsgroups: bionet.molbio.proteins Subject: Re: Proteins in Tears? Date: 7 Feb 1996 22:59:01 GMT Organization: Oklahoma State University, Stillwater OK Lines: 11 Message-ID: <4fbarl$ap9@news.cis.okstate.edu> References: <4f522v$qgn@netty.york.ac.uk> NNTP-Posting-Host: core.biochem.okstate.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) To: gst3@york.ac.uk X-URL: news:4f522v$qgn@netty.york.ac.uk Graham- Sorry I can't be of any help with specifics. I don't remember anything about who did the study or where published. But there was a study several (at least a dozen) years ago on components of human tears. They found an ingenious way to collect samples of subjects under emotional distress (gut-wrenching movie.) There were chemicals in these tears not found in the same subjects' samples collected under other conditions. They concluded that the therapeutic effect of tears has a biochemical basis. Good Luck From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!newsfeed.internetmci.com!in1.uu.net!news.ios.com!village.ios.com!kwas77 From: kwas77@village.ios.com (Robert Kwasnicki) Newsgroups: bionet.molbio.proteins Subject: Research by Angela Cacace - How to contact her? Date: 7 Feb 1996 22:44:26 GMT Organization: Internet Online Services Lines: 5 Message-ID: <4fba0a$2ag@news.ios.com> NNTP-Posting-Host: village.ios.com X-Newsreader: TIN [version 1.2 PL2] I am interested in contacting, via e-mail, phone, postal address, Dr. Angela M. Cacace. I believe she is now working at the NIH in Bethesda, MD. From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!Germany.EU.net!Frankfurt.Germany.EU.net!news.maz.net!news.isys.net!usenet From: janus.friedemann@hamburg.netsurf.de (Friedemann Janus) Newsgroups: bionet.molbio.proteins Subject: Are there Exonuclease-free bacterial strains ? Date: Thu, 08 Feb 1996 06:57:19 GMT Organization: iSYS Informationssystem Hamburg Lines: 11 Message-ID: <4fb3tc$14u@trance.isys.net> NNTP-Posting-Host: dip158-1.hamburg.netsurf.de X-Newsreader: Forte Free Agent 1.0.82 HI, I'm looking for bacterial strains which are deficient in overall exonuclease activity. In my project, I'd like to express proteins and screen for their exonuclease activity in a crude lysate. Any suggestions ? Friedemann From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!world!oravaxcm From: oravaxcm@world.std.com (Charles A Miller) Subject: How do you preflash x-rays? Message-ID: Organization: The World Public Access UNIX, Brookline, MA X-Newsreader: TIN [version 1.2 PL2] Date: Wed, 7 Feb 1996 19:06:58 GMT Lines: 10 I posted this before but I don't think it went through. How do you preflash x-ray film for Southern Blots? I'm doing ECL southern blotting and already have a camera flash. What kind of filter do I need and what are conditions (time, etc..) If anyone has any pointers I'd be most appreciative! Chuck oravaxcm@world.std.com From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!torn!nott!uottawa!bio02!ed From: Large Hearted Boy Newsgroups: bionet.molbio.proteins Subject: protein sequencing Date: Wed, 7 Feb 1996 21:47:32 -0500 Organization: University d'/of Ottawa Lines: 13 Message-ID: NNTP-Posting-Host: bio02.bio.uottawa.ca Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Hi everyone, I know that the reagents used in peptide sequencing are acetic anhydride and pyridine, acetic anhydride and aqueous NaCO3, ethanolic HCL, NBS, ninhydrin, Iodoacetic acid, performic acid... but where would these cleave ? how does the sequencing work ? thanks for any help you could give me. Ed Taboada Dept. of Biology University of Ottawa, Canada. From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Newsgroups: bionet.molbio.proteins,bionet.molbio.yeast,bionet.molbio.molec-model Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!gatech!newsfeed.internetmci.com!in1.uu.net!nih-csl!NewsWatcher!user From: newitt@nih.gov (John A. Newitt) Subject: Re: arrested nascent polypeptides Message-ID: Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: 128.231.113.35 Organization: National Institutes of Health X-Newsreader: Yet Another NewsWatcher 2.0.2 References: Date: Thu, 8 Feb 1996 01:42:13 GMT Lines: 32 Xref: biosci bionet.molbio.proteins:6972 bionet.molbio.yeast:4696 In article , meneghini@molbio.uoregon.edu (silverback) wrote: > o.k. does anyone know at which site (A or P) in the ribosome arrested > nascent chains lacking stop codons accumulate? Logic tells me that in the > absence of further message, the peptidyl tRNA may stay in the A site and > not translocate to the P site. However puromycin is capable of releasing > these nascent chains and the lore is (as I undersatnd it) that puromycin > only releases nascent chains in the P site. Please help, this information > is critical to my comprehensive exam proposals and no one in the > literature has offered any information on this. > thanks, > > marc meneghini > Institute of molecular biology, university of oregon I THINK that the terminal residue is in the P site and the puromycin sits in the A site. Puromycin enters at the A site, substituting for what would be an AA-tRNA following the terminal residue. See refs below for more info: "Ribosomes: Structure, Function, Genetics" G. Chamblis et al. (ed.) University Park Press, Baltimore (1980) Vazquez, D. (1979) "Molecular Biology Biochemistry and Biophysics" Vol. 30, Springer-Verlag, Berlin. Regards, John A. Newitt, Ph.D. | National Institutes of Health | FAX: 301-402-0387 Bethesda, Maryland USA | From owner-proteins@net.bio.net Tue Feb 06 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!news00.sunet.se!sunic!news99.sunet.se!news.uni-c.dk!news From: vadtsvet@biochem.ou.dk (Vadim Tsvetnitsky) Newsgroups: bionet.molbio.proteins Subject: Simulated HPLC of peptides Date: 7 Feb 1996 15:11:14 GMT Organization: Odense University, DENMARK Lines: 22 Message-ID: <4fafei$p8h@news.uni-c.dk> NNTP-Posting-Host: vadim.biochem.ou.dk X-Newsreader: WinVN 0.92.6+ Hi, I wonder if anybody knows if real-world peptide separation (order of elution from reverse-phase column) is different from software-simulated profile. I mean program calculates hydrophobicity of peptides or HPLC index and then predicts how they are going to be eluted from the standard RP-column. Any info is appreciated. |\/\/\/| | | | | | (o)(o) C _)_____________ | \___| / \ | / Be cool, man!| /____\ \_____________/ / \ Vadim Tsvetnitsky, Ph.D. vadtsvet@biochem.ou.dk ----------------------------------------------- From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!rutgers!concert!gatech!newsfeed.internetmci.com!uwm.edu!msunews!harbinger.cc.monash.edu.au!news.bhp.com.au!mel.dit.csiro.au!actcsiro!news.nsw.CSIRO.AU!metro!metro!inferno.mpx.com.au!news.unimelb.EDU.AU!news From: "Dr. Roger Murphy" Newsgroups: bionet.molbio.proteins Subject: Re: Tissue protein isolation? Date: Fri, 09 Feb 1996 14:51:16 +1000 Organization: University of Melbourne Lines: 40 Message-ID: <311AD2C4.265E@muwaye.unimelb.edu.au> References: <3117BF6B.2725@bmb-fs1.biochem.okstate.edu> NNTP-Posting-Host: murphy.pharmacology.unimelb.edu.au Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b6b (Win16; I) Ron Tate wrote: > > Jeffrey Upperman wrote: > > > > does anyone have a protocol or reference for tissue protein isolation? > > be gentle i am new to the protein world... > > -- > > Jeffrey S. Upperman,MD Internet: upperman@umdnj.edu > > Research Fellow Voice: (201) 982-4474 > > Dept. Anatomy, Cell Biology & Injury Fax: (201) 982-6803 > > Dept. of Surgery Beeper: (201) 708-4150 > > New Jersey Medical School Office: (201) 982-5045 > > 185 S. Orange Ave., G606 Lab: (201) 982-5404 > > Newark, NJ 07103 > > Jeffrey, > For starters check out "Protein Purification; Principals and Practice" > by Robert Scopes published by Springer-Verlag and volume 182 of the > "Methods in Enzymology" series. They both will give you guidelines > and techniques from tissue or cell disruption through almost any protein > purification technique you could want. > > Happy Hunting > > -- > Ron Tate > > Lab of Franklin Leach > Dept. of Biochem. & Molecular Biology > Oklahoma State University Try also "Protein purification applications" and "protein purification Methods", both by Harris and Angal, both in the "a practical approach" series from IRL Press - use 'em all the time myself! Roger Murphy Senior Research Fellow Pharmacology Department University of Melbourne From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!news.Stanford.EDU!nntp-hub2.barrnet.net!newsfeed.internetmci.com!uwm.edu!msunews!harbinger.cc.monash.edu.au!news.bhp.com.au!mel.dit.csiro.au!actcsiro!news.nsw.CSIRO.AU!metro!metro!inferno.mpx.com.au!news.unimelb.EDU.AU!news From: "Dr. Roger Murphy" Newsgroups: bionet.molbio.proteins Subject: Re: Simulated HPLC of peptides Date: Fri, 09 Feb 1996 14:56:55 +1000 Organization: University of Melbourne Lines: 35 Message-ID: <311AD417.13CE@muwaye.unimelb.edu.au> References: <4fafei$p8h@news.uni-c.dk> NNTP-Posting-Host: murphy.pharmacology.unimelb.edu.au Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b6b (Win16; I) Vadim Tsvetnitsky wrote: > > Hi, > > I wonder if anybody knows if real-world peptide separation (order of elution from reverse-phase > column) is different from software-simulated profile. I mean program calculates hydrophobicity > of peptides or HPLC index and then predicts how they are going to be eluted from the standard > RP-column. > > Any info is appreciated. > > |\/\/\/| > | | > | | > | (o)(o) > C _)_____________ > | \___| / \ > | / Be cool, man!| > /____\ \_____________/ > / \ > Vadim Tsvetnitsky, Ph.D. > vadtsvet@biochem.ou.dk > ----------------------------------------------- Hi Vadim! Depends on the size of the peptide - the larger they get, the more stable tertiary structure you see, so the more improbable the elution predictions become. For simple, small (<15 residues) peptides, elution prediction is reasonable relative to a standard. Absolute retention prediction is impossible because of the differences in selectivity of different adsorbants. Guess the motto is "suck it and see" - good luck! Roger Murphy From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!news.Stanford.EDU!nntp-hub2.barrnet.net!newsfeed.internetmci.com!primus.ac.net!news.cais.net!usenet.seri.re.kr!news.dacom.co.kr!nntp.coast.net!harbinger.cc.monash.edu.au!newshost.anu.edu.au!newsmaster From: Sven Johanson Newsgroups: bionet.molbio.proteins Subject: Immunoprecipitation from whole tissue Date: 9 Feb 1996 04:41:41 GMT Organization: Australian National University Lines: 15 Message-ID: <4feja5$34a@manuel.anu.edu.au> NNTP-Posting-Host: 150.203.193.93 Hi there! I've been trying to immunoprecipitate proteins from whole tissue (eg. nervous tissue) to look at protein associations with silver stained gels. As well as very weak signals I've had difficulties with heavy non-specific binding of proteins such as actin and others, using Dynabeads and Protein A-Seph. Preclearing and blocking with BSA don't appear to help. I don't know of anyone performing such experiments other than with cultured cells etc. and I'm beginning to understand why! Can anyone tell me more? Sven Johanson From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.tamu.edu!newshost.comco.com!news.texas.net!imci2!news.internetMCI.com!newsfeed.internetmci.com!in2.uu.net!world!oravaxcm From: oravaxcm@world.std.com (Charles A Miller) Subject: First: Having some trouble solubilizing a protein... Message-ID: Organization: The World Public Access UNIX, Brookline, MA X-Newsreader: TIN [version 1.2 PL2] Date: Fri, 9 Feb 1996 04:39:51 GMT Lines: 21 Hello, I am having trouble keeping a particular protein soluble. The protein is a HIS tagged fusion that I am purifying over a Nickel-NTA column under non-denaturing conditions. When the protein elutes, it is soluble for about 20 minutes, then begins to precipitate. It does the same thing in the presence of 50% glycerol. Some suggestions I have had include adding a sucrose solution into the cuvettes prior to elution so that they are eluted directly into the sugar solution. Before I crack open one of my pellets and begin, I would like to hear any feedback or suggestions. I have used a spreadsheet that I downloaded from the net that determines protein solubility. It tells me that the protein has a probability of being 60% insoluble (which is evident from its slow precipitation. By the way, it does not completely precipitate out of its elution buffer. It drops to a concentration of about 200ug/ml and then stops. If the precipitate is spun out and the supernatant removed, no more protein "drops" out. Hope someone can give me a few pointers or even a few references... Chuck Miller oravaxcm@world.std.com From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsserver.jvnc.net!netnews.sb.com!news From: Lynne Donegan Newsgroups: bionet.molbio.proteins Subject: Re: gfp sensitivity Date: 8 Feb 1996 16:28:11 GMT Organization: SmithKline Beecham Lines: 25 Message-ID: <4fd8ar$cc5@hbu005.ha.uk.sbphrd.com> References: NNTP-Posting-Host: had938.ha.uk.sbphrd.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) bz182@FREENET.TORONTO.ON.CA (Wei Hu) wrote: >hi there, >i don't know whether gfp can be used to trace a low abundance protein in >mammalian cells, which is usually pulse-labeled with 35s-met. >could anyone experienced in gfp please tell me the related information. >many thanks in advance. >wei > Personally, I have no experience with gfp although I am planning to get my hands on it soon. There is a news group called bionet.molbiol.proteins.flourescent (I think thats it anyway!) that you may find helpful Just an idea Lynne -- The opinions expressed in this communication are my own, and do not necessarily reflect those of my employer. From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!GUARANY.CPD.UNB.BR!zanotta From: zanotta@GUARANY.CPD.UNB.BR (pedro jose portugal zanotta) Newsgroups: bionet.molbio.proteins Subject: Re: Measuring Peptide Concentration Date: 8 Feb 1996 08:39:59 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 28 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <4f81j1$1m6@griffin.ccc.nottingham.ac.uk> NNTP-Posting-Host: net.bio.net On 6 Feb 1996, Allister Maynard wrote: > I have a problem measuring the concentraion of a micromolar solution of a > peptide. It is ubiquitin derived, 16 amino acids in length. I do not have > enough to accurately weigh so I have dissolved it in water to try UV. I know > the sequence but all the quantitation methods I have found so far rely on the > peptide having tryosine, histidine or cystine residues, all of which are absent > in my sequence. > > Is there a way to reasonably accurately find the concentration of my peptide > solution in a non-destructive way? > > Many thanks to all who reply. > _______________________________________________________________________ > | | > | ,_ o Al Maynard, | > | / //\, B9a Chemistry, University of Nottingham. | > | \>> | Phrase of the now:'An Idea a day keeps the supervisor away!`| > | \\, E-mail: pcxam@unix.ccc.nottingham.ac.uk | > |_______________________________________________________________________| > Maynard, You could try read the absortion in the 215-216 nm which is where the peptide bond have its maximum of absorbance. Good luck! Pedro Zanotta> > From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.dacom.co.kr!nntp.coast.net!col.hp.com!news.cis.okstate.edu!usenet From: Ron Tate Newsgroups: bionet.molbio.proteins Subject: Re: Measuring Peptide Concentration Date: Thu, 08 Feb 1996 09:53:04 -0600 Organization: Dept. of Biochemistry and Molecular Biology, OSU Lines: 34 Message-ID: <311A1C60.3982@bmb-fs1.biochem.okstate.edu> References: <4f81j1$1m6@griffin.ccc.nottingham.ac.uk> NNTP-Posting-Host: firefly3.biochem.okstate.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b5 (Win95; I) Allister Maynard wrote: > > I have a problem measuring the concentraion of a micromolar solution of a > peptide. It is ubiquitin derived, 16 amino acids in length. I do not have > enough to accurately weigh so I have dissolved it in water to try UV. I know > the sequence but all the quantitation methods I have found so far rely on the > peptide having tryosine, histidine or cystine residues, all of which are absent > in my sequence. > > Is there a way to reasonably accurately find the concentration of my peptide > solution in a non-destructive way? > > Many thanks to all who reply. > _______________________________________________________________________> | | > | ,_ o Al Maynard, | > | / //\, B9a Chemistry, University of Nottingham. | > | \>> | Phrase of the now:'An Idea a day keeps the supervisor away!`| > | \\, E-mail: pcxam@unix.ccc.nottingham.ac.uk | > |_______________________________________________________________________| Al, Try absorbance at 230 and 260nm, it is reported to be as sensitive as the Lowry method but with much less protein to protein variability. The concentration equation is microgram/ml= 183 x A230 - 75.8 x A260. The reference is "Analytical Biochemistry" volume 82 pp. 362-371. 1977. Hope it helps. Happy Hunting -- Ron Tate Lab of Franklin Leach Dept. of Biochem. & Molecular Biology Oklahoma State University From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!news00.sunet.se!sunic!news99.sunet.se!news.uni-c.dk!eagle.novo.dk!usenet From: "Jack B. Nielsen" Newsgroups: bionet.molbio.proteins Subject: Re: Proteins in Tears? Date: 8 Feb 1996 12:24:10 GMT Organization: Novo Nordisk A/S Lines: 32 Message-ID: <4fcq1a$dp7@eagle.novo.dk> References: <4f522v$qgn@netty.york.ac.uk> NNTP-Posting-Host: jabn.novo.dk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) To: gst3@york.ac.uk Below are proteins identified from humans. You may also find interesting data in McGill, 1984 "British journal of opthalmology" Best reagards, Jack Gachon, 1983 Chao, 1987 Coyle,1989 Albumin + + + 1 antichymotrypsin + + 1 antitrypsin + + ceruloplasmin + Zn 2 glycoprotein + + 1-acid glycoprotein + haptoglobin + IgA -chain + + IgA + + IgE + IgG + + IgM + lysozyme + + + 2-macroglobulin + + secretory component + + + transferrin + + + lactoferrin. + + + PMFA + + Table: Proteins identified in tears or human ocular mucus. From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!usenet.eel.ufl.edu!warwick!news.nott.ac.uk!griffin.nott.ac.uk!usenet From: Allister Maynard Newsgroups: bionet.molbio.proteins Subject: Measuring Peptide Concentration Date: 6 Feb 1996 17:02:25 GMT Organization: Cripps Computing Centre, The University of Nottingham Lines: 19 Message-ID: <4f81j1$1m6@griffin.ccc.nottingham.ac.uk> NNTP-Posting-Host: ccsun7.ccc.nottingham.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12 (X11; I; SunOS 5.5 sun4c) X-URL: news:bionet.molbio.proteins I have a problem measuring the concentraion of a micromolar solution of a peptide. It is ubiquitin derived, 16 amino acids in length. I do not have enough to accurately weigh so I have dissolved it in water to try UV. I know the sequence but all the quantitation methods I have found so far rely on the peptide having tryosine, histidine or cystine residues, all of which are absent in my sequence. Is there a way to reasonably accurately find the concentration of my peptide solution in a non-destructive way? Many thanks to all who reply. _______________________________________________________________________ | | | ,_ o Al Maynard, | | / //\, B9a Chemistry, University of Nottingham. | | \>> | Phrase of the now:'An Idea a day keeps the supervisor away!`| | \\, E-mail: pcxam@unix.ccc.nottingham.ac.uk | |_______________________________________________________________________| From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsserver.jvnc.net!news.Edu.TW!news.cc.nctu.edu.tw!nctuccca.edu.tw!netnews.ntu.edu.tw!news From: "C. S. Liu" Newsgroups: bionet.molbio.proteins Subject: HELP - Aminopeptidase M Date: 8 Feb 1996 03:53:43 GMT Organization: National Taiwan University Lines: 7 Message-ID: <4fbs47$3t5@netnews.ntu.edu.tw> NNTP-Posting-Host: @140.112.72.182 Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Mailer: Mozilla 1.2N (Windows; I; 16bit) Does anyone have the experience of using an aminopeptidase M to obtain N-terminally truncated glycoprotein like SY¡ôLPPC......? Please give me a detailed experimental account. Thank you very much. From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!news.Stanford.EDU!nntp-hub2.barrnet.net!news1.digital.com!ames!waikato!auckland.ac.nz!sbs-43-129.sbs.auckland.ac.nz!user From: humanmol@auckland.ac.nz (Human Mol genetics) Newsgroups: bionet.molbio.proteins Subject: Stoke's radius Date: Fri, 09 Feb 1996 16:11:35 +1300 Organization: e Lines: 24 Message-ID: NNTP-Posting-Host: sbs-43-129.sbs.auckland.ac.nz Can anyone direct me to a source of values for the Stoke's radius of the following; BSA Ovalbumin Carbonic anhydrase Bovine ribonuclease Aprotinin Thank You Liam Williams and John Taylor -- --------------------------------------------------------------------- Andrew Dodd Molecular Genetics and Microbiology School of Biological Sciences University of Auckland Private Bag 92019 Phone 64-9-3737599 x7232 Auckland Fax: 64-9-3737414 NEW ZEALAND E-mail: a.dodd@auckland.ac.nz --------------------------------------------------------------------- From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!CMU.CHIANGMAI.AC.TH!mdbci001 From: mdbci001@CMU.CHIANGMAI.AC.TH (Prachya Kongtawelert) Newsgroups: bionet.molbio.proteins Subject: Re: Is there any limitation of using BioRad method to..? Date: 8 Feb 1996 18:35:36 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <4fdlc2$c61$1@mhade.production.compuserve.com> NNTP-Posting-Host: net.bio.net When working with BCA assay, plese be careful if your sample is biotinylated form or contaminated with biotin. It make false positive and give you over estimate of protein. Prachya On 8 Feb 1996, Geoffrey D. Wheelock wrote: > you don't want to use the Bradford assay when detergents are present > (so maybe Na-Cholate is the problem). Try Pierce's BCA assay > instead. Good luck > > From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bloom-beacon.mit.edu!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!news00.sunet.se!sunic!news99.sunet.se!umdac!news From: "Leszek A. Kleczkowski" Newsgroups: bionet.molbio.proteins Subject: the smallest kinase? Date: 8 Feb 1996 23:46:31 GMT Organization: University of Umea, Sweden Lines: 16 Message-ID: <4fe20n$rd1@studium.student.umu.se> NNTP-Posting-Host: leszek.fysbot.umu.se Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) > MessageID: kklee-2701961422130001@virus23.virus.kyoto-u.ac.jp#1/1 > Dose anyone knows the smallest kinase ? > I think cdk may be the smallest ranging in molecular weight around KDa. > I am purifing a kinase activity of ~34KDa, which may not be cdk family. > So, I want to any information from everyone of the smallest kinase. There is adenylate kinase which has been quoted to have ca. 20-27 kd, depending on species, in both plant and animal tissues. If I remember correctly it is also the only kinase which uses free (no magnesium) nucleotide as one of the reactants. Leszek From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!usenet.eel.ufl.edu!usenet.cis.ufl.edu!eng.ufl.edu!clas.ufl.edu!mailey!susanbr From: Susan Rasmussen Newsgroups: bionet.molbio.proteins Subject: Metal Affinity Purification Date: Thu, 8 Feb 1996 15:58:53 -0500 Organization: University of Florida's College of Liberal Arts and Sciences Lines: 10 Message-ID: NNTP-Posting-Host: server.chem.ufl.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Using Nickel affinity purification columns, I am experiencing losses in activity in my protein. This is due to the loss of disulfide bonds in the protein. My question is if anyone has experienced the loss of disulfide bonds while using this method of purification. And if so, what was done to overcome this problem. Thanks for your help. Susan B. Rasmussen From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.compuserve.com!news.production.compuserve.com!news From: Geoffrey D. Wheelock <74710.2416@CompuServe.COM> Newsgroups: bionet.molbio.proteins Subject: Re: Is there any limitation of using BioRad method to..? Date: 8 Feb 1996 20:10:42 GMT Organization: Paracelsian, Inc. Lines: 3 Message-ID: <4fdlc2$c61$1@mhade.production.compuserve.com> References: <4fa9u6$o2d@hpg30a.csc.cuhk.hk> you don't want to use the Bradford assay when detergents are present (so maybe Na-Cholate is the problem). Try Pierce's BCA assay instead. Good luck From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!newsfeed.pitt.edu!quadra950.pathology.pitt.edu!user From: mcgorry@med.pitt.edu (Michael C. Gorry) Newsgroups: bionet.molbio.methds-reagnts,bionet.cellbiol,bionet.microbiology,bionet.molbio.proteins,sci.bio.microbiology Subject: bacterial housekeeping genes Date: 8 Feb 1996 18:36:40 GMT Organization: Univ of Pittsburgh, Pathology Lines: 14 Message-ID: NNTP-Posting-Host: quadra950.pathology.pitt.edu Xref: biosci bionet.molbio.methds-reagnts:40017 bionet.cellbiol:3990 bionet.microbiology:4894 bionet.molbio.proteins:6981 sci.bio.microbiology:2624 I have been looking for a bacterial housekeeping gene that has a high level of expression. Would GADPH be suitable? We are looking for any gene that would have a high mRNA level within a cell. I haven't been able to find anything published that discusses expression rates within cells. There was, however, a mention of using GADPH for an RT-PCR quantitative control. Is anyone aware of other "controls" that have been used to quantitate PCR reactions? I would appreciate any advice. Thanks -- Michael Gorry mcgorry@med.pitt.edu From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Path: biosci!JASON.UTHCT.EDU!SHAUN From: SHAUN@JASON.UTHCT.EDU ("Shaun D. Black") Newsgroups: bionet.molbio.proteins Subject: Re: Measuring Peptide Concentration Date: 8 Feb 1996 09:30:20 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 14 Sender: daemon@net.bio.net Distribution: world Message-ID: <960208113035.9a8f@jason.uthct.edu> NNTP-Posting-Host: net.bio.net Hi, You might try the Abs 205 nm method to determine the concentration of your peptide. I've done this many times, and it works just fine, notably because it is almost solely based on peptide-bond absorbance, versus side- chains (Tyr, Cys, etc.). Check out Methods Enzymol. vol. 182 "Quantitation of Protein" by Christa Stoscheck (1990), and see the section beginning on page 55 specially. I hope this helps you a bit. Best regards, Shaun =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= = Shaun D. Black, PhD | Internet address: shaun@jason.uthct.edu = = Dept. of Biochemistry | University of Texas Health Center, at Tyler = = World Wide Web: http://pegasus.uthct.edu/UTHCT-Home/Welcome.html = =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= From owner-proteins@net.bio.net Wed Feb 07 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!rutgers!csn!magnus.acs.ohio-state.edu!lerc.nasa.gov!purdue!gatech!newsfeed.internetmci.com!in2.uu.net!world!oravaxcm From: oravaxcm@world.std.com (Charles A Miller) Subject: Second: Need references for hydropathy and structure determination.. Message-ID: Organization: The World Public Access UNIX, Brookline, MA X-Newsreader: TIN [version 1.2 PL2] Date: Fri, 9 Feb 1996 04:45:37 GMT Lines: 15 Hello again, I have been using a couple programs to determine Enzyme cutting points in DNA as well as Hydropathy. Unfortunately, I do not have a good background in analyzing hydropathy plots or alpha-helix/beta-sheet plots. I would like to be able to determine or at least make an educated hypothesis as to the location of trans membrane domains and outer/inner membrane domains of proteins using hydropathy and helix/sheet presence. If anyone has any good suggestions for references (books or articles or programs) regarding this subject, I'd be most grateful. Thanks a lot! Chuck Miller oravaxcm@world.std.com From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!KB.USM.MY!microlab From: microlab@KB.USM.MY (Makmal Mikrobiologi) Newsgroups: bionet.molbio.proteins Subject: unsubscribe Date: 9 Feb 1996 01:31:58 -0800 Organization: Universiti Sains Malaysia Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net unsubscribe protiens@net.bio.net From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!rutgers!gatech!usenet.eel.ufl.edu!psgrain!news.hklink.net!news.asiaonline.net!NewsWatcher!user From: sansha@asiaonline.net (Jonathan Robson) Newsgroups: bionet.molbio.proteins Subject: ovoflavoprotein sequence in hen's egg Date: 10 Feb 1996 03:17:39 GMT Organization: Sansha Productions Ltd Lines: 12 Message-ID: NNTP-Posting-Host: ip34.asiaonline.net My friend Prof. Rao Ping Fan at Fuzhou university in China is searching for the amino acid sequence to hens egg ovoflavoprotein. As the entire university is only allowed 4 hrs per day online it is taking a while to process his request so I am trying to find it from my office in Hong Kong. I have already searched several databases for him but no luck so far although as I am not a biochemist it is possible I missed it. If anyone can help, please send me mail to sansha@asiaonline.net -- Jonathan Robson Sansha Productions Ltd http://sansha.com/sansha From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!agate!howland.reston.ans.net!newsfeed.internetmci.com!primus.ac.net!news.cais.net!usenet.seri.re.kr!news.kreonet.re.kr!chiak.kaist.ac.kr!dykim From: dykim@chiak.kaist.ac.kr (KIm doo yeon) Newsgroups: bionet.molbio.proteins Subject: Need Fv fragment to phoP-Tyr Date: 10 Feb 1996 03:16:01 GMT Organization: Korea Research Environment Open Network (KREONet) Lines: 19 Message-ID: <4fh2lh$76i@news.kreonet.re.kr> NNTP-Posting-Host: chiak.kaist.ac.kr X-Newsreader: TIN [version 1.1 PL9] Hi, I need the Fv fragment genes encoding the Fv fragment recognizing the phosphotyrosine, phosphoserine or phosphothreonine. Where can I get these genes? Any information regarding the possibility of getting these genes will be greatly appreciated. Please Help Me ! __________________________________________________________________________ KAIST, Deajon Korea. Dooyeon Kim ...................... dykim@bioneer.kaist.ac.kr ___________________________________________________________________________ From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!newsserver.jvnc.net!news.join.ad.jp!news.imnet.ad.jp!ripspost.aist.go.jp!news.tisn.ad.jp!tkyex1.phys.s.u-tokyo.ac.jp!news.nc.u-tokyo.ac.jp!t-server!mech.t.u-tokyo.ac.jp!130.69.160.103!yokada From: yokada@kinesin.kaibo1.m.u-tokyo.ac.jp (Yasushi OKADA) Newsgroups: bionet.molbio.proteins Subject: Re: Metal Affinity Purification Date: 9 Feb 1996 13:43:36 GMT Organization: Dept. Anatomy & Cell Biol. Facul. Med. Univ. Tokyo. Lines: 33 Message-ID: References: NNTP-Posting-Host: kinesin.kaibo1.m.u-tokyo.ac.jp In-reply-to: Susan Rasmussen's message of Thu, 8 Feb 1996 15:58:53 -0500 My protein is also affected by metal affinity purification probably due to the oxidation of cys-residue. Part of metal ion (Ni++ or Co++) is reduced (becomes Ni or Co) when incubated with my protein. Then, my protein loses its activity. I have found that the addition of BSA and/or Triton X-100 (non-oxidized, membrane protein purification grade), use of minimal amount of column and the removal of oxigen from from solutions can partly inhibit this effect. My protein is expressed with insect cells (Sf21). I also found that the carry over of the culture medium to the cell homogenate enhances this effect. Yasushi Okada. >>>>> On Thu, 8 Feb 1996 15:58:53 -0500, Susan Rasmussen said: > Using Nickel affinity purification columns, I am experiencing > losses in activity in my protein. This is due to the loss of > disulfide bonds in the protein. My question is if anyone has > experienced the loss of disulfide bonds while using this method of > purification. And if so, what was done to overcome this problem. > Thanks for your help. > Susan B. Rasmussen -- ****************************************************************************** * Yasushi Okada, MD. | Email:yokada@kinesin.kaibo1.m.u-tokyo.ac.jp * * Dept. Anatomy & Cell Biology | Tel:81-3-3812-2111 ext.3336 * * Univ. Tokyo JAPAN | Fax:81-3-5689-4856 * ****************************************************************************** From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!rutgers!gatech!newsfeed.internetmci.com!news.sprintlink.net!news.voicenet.com!ivyland250.voicenet.com!gheavner From: gheavner@voicenet.com (George A. Heavner) Newsgroups: bionet.molbio.proteins Subject: Re: Simulated HPLC of peptides Date: Fri, 9 Feb 1996 17:22:19 LOCAL Organization: Voicenet - Internet Access - (215)674-9290 Lines: 36 Message-ID: References: <4fafei$p8h@news.uni-c.dk> <311AD417.13CE@muwaye.unimelb.edu.au> NNTP-Posting-Host: ivyland250.voicenet.com X-Newsreader: Trumpet for Windows [Version 1.0 Rev B final beta #4] I don't know how the field has progressed in the last few years. I can recall reading a paper that claimed to be able to predict elution times based on composition and sequence. At the time I was working with a small peptide that, according to the calculations, should have eluted 2-3 minutes before being injected. George A. Heavner Senior Director, Peptide R&D Centocor, Inc. +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ >> Hi, >> >> I wonder if anybody knows if real-world peptide separation (order of elution from reverse-phase >> column) is different from software-simulated profile. I mean program calculates hydrophobicity >> of peptides or HPLC index and then predicts how they are going to be eluted from the standard >> RP-column. >> >> Any info is appreciated. >> >> |\/\/\/| >> | | >> | | >> | (o)(o) >> C _)_____________ >> | \___| / \ >> | / Be cool, man!| >> /____\ \_____________/ >> / \ >> Vadim Tsvetnitsky, Ph.D. >> vadtsvet@biochem.ou.dk >> ----------------------------------------------- From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!CS.Arizona.EDU!news.Arizona.EDU!hamblin.math.byu.edu!park.uvsc.edu!news.caldera.com!news.cc.utah.edu!news From: zinc@zifi.genetics.utah.edu (zinc) Newsgroups: bionet.molbio.proteins Subject: Re: First: Having some trouble solubilizing a protein... Date: 9 Feb 1996 17:40:48 GMT Organization: University of Utah Computer Center Lines: 57 Message-ID: <4fg0v0$bp0@news.cc.utah.edu> References: NNTP-Posting-Host: zifi.genetics.utah.edu X-Newsreader: knews 0.9.5 In-Reply-To: To: oravaxcm@world.std.com (Charles A Miller) -----BEGIN PGP SIGNED MESSAGE----- In article , oravaxcm@world.std.com (Charles A Miller) writes: >Hello, > I am having trouble keeping a particular protein soluble. The protein >is a HIS tagged fusion that I am purifying over a Nickel-NTA column under >non-denaturing conditions. When the protein elutes, it is soluble for >about 20 minutes, then begins to precipitate. It does the same thing in the >presence of 50% glycerol. Some suggestions I have had include adding a sucrose >solution into the cuvettes prior to elution so that they are eluted directly >into the sugar solution. Before I crack open one of my pellets and begin, >I would like to hear any feedback or suggestions. I have used a >spreadsheet that I downloaded from the net that determines protein >solubility. It tells me that the protein has a probability of being >60% insoluble (which is evident from its slow precipitation. By the way, it >does not completely precipitate out of its elution buffer. It drops to a >concentration of about 200ug/ml and then stops. If the precipitate is spun >out and the supernatant removed, no more protein "drops" out. > >Hope someone can give me a few pointers or even a few references... Chuck, i've seen this sort of thing as well with my protein. i think the problem has two parts. the first is that the Ni++ affinity column has an extremely high capacity for protein which allows you to load quite a lot of extract on the column. the second part is that if you elute by just adding the high imidazole buffer you essentially remove all of the protein at once. so now you've got a lot of protein that's fairly concentrated in a high salt buffer (~1M NaCl if i recall). this is asking for the protein to agregate due to the hydrophobic effect. i suppose the solution is to just load less protein or to elute with a linear gradient such that the protein is not so concentrated. - -pjf - -- "Those that give up essential liberty to obtain a little temporary safety deserve neither liberty nor safety." -- Benjamin Franklin (1773) finger for PGP key zifi runs LINUX 1.3.57 -=-=-=WEB=-=-=-> http://zifi.genetics.utah.edu -----BEGIN PGP SIGNATURE----- Version: 2.6.2 Comment: Say NO to GAK! - fight for your privacy! iQCVAwUBMRuHRU3Qo/lG0AH5AQGMhQP/dbgQ3ckNH7yL0s92GHmHVbN1oaepxevc FsfRneJh7jat6wu25DJlP0wawDt1IIDbEDbNUEtZfoMWLdUwJPNM9sIRU+RoNYU3 Q7r7/C54tUjphILZcpPOgJxOicoLT+N48wA8OKTMkvhKTdGXOwBTZ2F6eWiJkQIM D5vl2R5lyS8= =J3lO -----END PGP SIGNATURE----- From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!nntp.coast.net!swidir.switch.ch!scsing.switch.ch!news.belwue.de!fu-berlin.de!cs.tu-berlin.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins Subject: Re: Is there any limitation of using BioRad method to..? Date: 8 Feb 1996 11:13:05 GMT Organization: University of Wuerzburg, Germany Lines: 34 Message-ID: <4fcls1$s9r@winx03.informatik.uni-wuerzburg.de> References: <4f7bu5$9ls@hpg30a.csc.cuhk.hk> <4fa9u6$o2d@hpg30a.csc.cuhk.hk> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Ricky Leung (s935537@acs.csc.cuhk.hk) wrote: > All suggestions are welcome > One additional information I have to write is the sample was dissolved > in potassium chloride and sodium cholate. > Was the precipitation related to sodium cholate? That is indeed possible. The following agents give changes in the Bradford assay (list from Bradford's Anal. Biochem. paper, is certainly not exhaustive): Agent approx. OD change 2 M Tris 0.026 99% Glycerol 0.012 1 M Sucrose 0.013 Acetone 0.069 5% Phenol 0.046 0.1% Triton X-100 0.013 1% Triton X-100 0.590 1% SDS 0.495 1% Haemosol 0.108 I have experienced that 500 mM potassium phosphate pH 6.8 gives an OD change of larger than 2 in the micro-assay. I suspect this is due to a pH change in the solution since this high concentration will probably buffer the phosphoric acid which is used to dissolve the CBB in. However, I have been too lazy to do exhaustive tests. --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!lhc.nlm.nih.gov!crick.sura.net!lamarck.sura.net!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!torn!nott!nrcnet0.nrc.ca!BRI.NRC.CA!Feng.Ni From: Feng.Ni@BRI.NRC.CA (Feng Ni) Newsgroups: bionet.molbio.proteins Subject: POSITIONS IN PROTEIN NMR - SECOND ANOUNCEMENT Date: 9 Feb 1996 18:56:13 GMT Organization: Institut de recherche en biotechnologie, Montréal Lines: 39 Distribution: world Message-ID: <4fg5cd$766@nrcnet0.nrc.ca> NNTP-Posting-Host: indy.bri.nrc.ca POSTDOCTORAL AND RESEARCHER POSITIONS IN BIOPHYSICAL AND PROTEIN NMR SPECTROSCOPY Postdoctoral and researcher positions are available in the NMR laboratory of the Biotechnology Research Institute (BRI). Our research centers on proteins in blood coagulation, protease regulation and ligand-receptor interactions. Candidates must have a recent Ph.D. degree in chemistry, biochemistry or a related field and have demonstrated abilities (e.g. through publications) in carrying out creative research. Knowledge of high-resolution NMR spectroscopy is desirable and experience in protein/peptide purification is an important asset. Through their work at BRI, postdoctoral fellows obtain experience in the use of NMR spectroscopy to determine the structures and the biophysical properties of proteins and protein-protein complexes. Postdoctoral fellows are paid an annual salary of $23,000-$28,000 according to the MRC guidelines. Exceptional candidates may be promoted to senior research positions offerred by the National Research Council of Canada. Our institute is equipped with one Bruker AMX-500 NMR spectrometer and shares a second with members of the Montreal Joint Centre for Structural Biology. Funds are secured for a state-of-the-art higher field NMR instrument to be installed within the coming months. Qualified individuals are invited to forward your resume to: Feng Ni, Ph.D. Head of Biomolecular NMR Biotechnology Research Institute 6100 Royalmount Avenue Montreal, Quebec, H4P 2R2 Canada phone: (514)-496-6729 fax: (514)-496-5143 e_mail: fengni@bri.nrc.ca Applications will be accepted until the positions are filled. We thank all applicants for their interests, however, only those being considered will be contacted. From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!CS.Arizona.EDU!news.Arizona.EDU!hamblin.math.byu.edu!acs2.byu.edu!news.cuny.edu!ukma!gatech!swrinde!cssun.mathcs.emory.edu!hobbes.cc.uga.edu!mac200.ccrc.uga.edu!user From: rmerkle@uga.cc.uga.edu (Roberta K. Merkle) Newsgroups: bionet.molbio.proteins Subject: Laboratory Courses - Characterization of Complex Carbohydrates Date: Fri, 09 Feb 1996 11:48:11 -0500 Organization: CCRC - University of Georgia Lines: 32 Message-ID: NNTP-Posting-Host: mac200.ccrc.uga.edu Techniques for Characterization of Complex Carbohydrates Laboratory Courses Three courses will be offered at the Complex Carbohydrate Research Center (CCRC) of the University of Georgia. Course 1 (June 10-14, 1996), "Separation and Characterization of Glycoprotein Oligosaccharides", is intended for scientists with little experience in carbohydrate analysis. Course 2 (June 17-21, 1996), "Structural Analysis of Oligosaccharides", is intended for scientists with some experience with glycoconjugates or for those who have completed the first course, and will focus on techniques of composition and linkage analysis. Experience with basic biochemical techniques is a prerequisite for participation in the courses. Course 3 (August 19-23, 1996) "MS and MS/MS Analysis of Glycoconjugates" is intended for scientists with an interest in mass spectrometry and some experience with glycoconjugates. Experience with MS is beneficial but is not required. All courses will consist of hands-on laboratory work, demonstrations and lectures; lab manual including selected analytical techniques and references will be provided. Each course is limited to 10 participants. The cost of registration (including lunch) per course is $500 for individuals from non-profit institutions, $1,100 for others. Registration does not include lodging and travel expenses. The courses are supported jointly by the Department of Energy Plant and Microbial Carbohydrate Center, and the NIH Biomedical Carbohydrate Resource Center of the CCRC. For further information or to apply for the courses contact: Dr. Roberta K. Merkle, Complex Carbohydrate Research Center, 220 Riverbend Road, The University of Georgia, Athens, Georgia 30602-4712. Phone: 706-542-4402. Facsimile: 706-542-4412. E-mail: RMERKLE@UGA.CC.UGA.EDU From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!bloom-beacon.mit.edu!newsfeed.internetmci.com!usenet.eel.ufl.edu!usenet.cis.ufl.edu!usenet.ufl.edu!biomac1.health.ufl.edu!user From: regina@biotech.ufl.edu (Regina Shaw) Newsgroups: bionet.molbio.proteins Subject: detergents in ELISA Date: 9 Feb 1996 19:12:44 GMT Organization: University of Florida Lines: 3 Message-ID: NNTP-Posting-Host: biomac1.health.ufl.edu To everyone who responded to my message ("do detergents interfere in ELISA"): Thank you very much for all your helpful comments! Regina From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!lhc.nlm.nih.gov!crick.sura.net!lamarck.sura.net!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!torn!utnut!oci!news From: Flip Hoedemaeker Subject: Re: Simulated HPLC of peptides Content-Type: text/plain; charset=us-ascii Message-ID: <311B6285.2781@oci.utoronto.ca> Sender: news@oci.utoronto.ca Content-Transfer-Encoding: 7bit Organization: Ontario Cancer Institute - U of Toronto References: <4fafei$p8h@news.uni-c.dk> <311AD417.13CE@muwaye.unimelb.edu.au> Mime-Version: 1.0 Date: Fri, 9 Feb 1996 15:04:37 GMT X-Mailer: Mozilla 2.0 (X11; I; IRIX 5.3 IP20) Lines: 45 Dr. Roger Murphy wrote: > > Vadim Tsvetnitsky wrote: > > > > Hi, > > > > I wonder if anybody knows if real-world peptide separation (order of elution from reverse-phase > > column) is different from software-simulated profile. I mean program calculates hydrophobicity > > of peptides or HPLC index and then predicts how they are going to be eluted from the standard > > RP-column. > > > > Any info is appreciated. > > > > |\/\/\/| > > | | > > | | > > | (o)(o) > > C _)_____________ > > | \___| / \ > > | / Be cool, man!| > > /____\ \_____________/ > > / \ > > Vadim Tsvetnitsky, Ph.D. > > vadtsvet@biochem.ou.dk > > ----------------------------------------------- > > Hi Vadim! > > Depends on the size of the peptide - the larger they get, the more stable > tertiary structure you see, so the more improbable the elution > predictions become. For simple, small (<15 residues) peptides, elution > prediction is reasonable relative to a standard. Absolute retention > prediction is impossible because of the differences in selectivity of > different adsorbants. Guess the motto is "suck it and see" - good luck! > > Roger Murphy To add to this, I did some purification of complex peptide mixtures on RF HPLC (trypsin digests of proteins) and I have noticed that elution of one peptide can prematurely trigger (partial) elution of another. You'll see the latter peptide in three of four different peaks sometimes. I guess that if you use large concentrations, peptides not only bind directly to the matrix but also to each other. Flip From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!gatech!usenet.eel.ufl.edu!warwick!news.nott.ac.uk!griffin.nott.ac.uk!usenet From: Keith Bradnam Newsgroups: bionet.general,bionet.celegans,bionet.molbio.proteins Subject: Seeking info on cpr genes Date: 8 Feb 1996 12:03:32 GMT Organization: Department of Genetics Lines: 15 Message-ID: <4fcoqk$c5j@griffin.ccc.nottingham.ac.uk> NNTP-Posting-Host: evol.gene.nottingham.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (X11; I; OSF1 V3.2 alpha) X-URL: news:bionet.general Xref: biosci bionet.general:19836 bionet.celegans:800 bionet.molbio.proteins:6996 In C.elegans there are 4 genes (cpr-3, cpr-4, cpr-5 and cpr-6) that encode a cathepsin B-like cysteine proteinase. I'm looking for more info on these genes/proteins as the original paper cited in the GenBank entry provided no information at all. Does anyone know about the function and expression patterns of these genes? If so I would be very grateful if you could provide me with either information or suitable references. Yours hopefully, Keith Bradnam From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!agate!howland.reston.ans.net!newsfeed.internetmci.com!primus.ac.net!news.cais.net!xara.net!zynet.net!bsdi002.britain.eu.net!uknet!ftel.co.uk!bham!usenet From: altabios@bham.ac.uk (John E. Fox) Newsgroups: bionet.molbio.proteins Subject: Re: protein sequencing Date: 9 Feb 1996 13:19:05 GMT Organization: Alta Bioscience Lines: 15 Message-ID: <4ffhk9$c2i@sun4.bham.ac.uk> References: NNTP-Posting-Host: bcs118.bham.ac.uk X-Newsreader: WinVN 0.90.6 In article , Large Hearted Boy says: > >Hi everyone, > >I know that the reagents used in peptide sequencing >are acetic anhydride and pyridine, acetic anhydride and aqueous NaCO3, >ethanolic HCL, NBS, ninhydrin, Iodoacetic acid, performic acid... Sorry, you've been mis-informed, none of these are used in sequencing. Try 'Protein Sequencing a practical approach' ed J.B.C.Findley M.J. Geisow IRL press 1989 John Fox From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!daresbury!bioftp.unibas.ch!infobiogen.fr!pasteur.fr!jussieu.fr!univ-lyon1.fr!news.imag.fr!ciril.fr!usenet From: Clarisse Perrin Newsgroups: bionet.molbio.proteins Subject: Postranslational changes in bacteria? Date: 9 Feb 1996 10:15:01 GMT Organization: Universite Henry Poincare Lines: 12 Message-ID: <4ff6r5$jkj@arcturus.ciril.fr> NNTP-Posting-Host: debussy.scbiol.u-nancy.fr Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (X11; I; SunOS 4.1.3C sun4m) X-URL: news://arcturus.ciril.fr/bionet.molbio.proteins We have purified a 16 kDa protein from streptococcus. In HPLC C4 there are four pics with retentions times of 12, 17, 22 and 44 min. It seems that the N-terminus sequence of four proteins its the same. This change in RT could be due to postranslational changes in protein. Any one has an idea. Thanks for help. Clarisse Perrin Food Biosciences Henri Poincare University 54506 Vandoeuvre-les-Nancy France From owner-proteins@net.bio.net Thu Feb 08 22:00:00 1996 Path: biosci!rutgers!gatech!usenet.eel.ufl.edu!cobia.gulf.net!news.duke.edu!bio1.acpub.duke.edu!mmc9 From: Mara Michelle Casar Newsgroups: bionet.molbio.proteins Subject: receptor binding - help! Date: Fri, 9 Feb 1996 14:34:55 -0500 Organization: Duke University, Durham, NC, USA Lines: 16 Message-ID: NNTP-Posting-Host: bio1.acpub.duke.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII I'm having a bit of trouble with something I'm working on right now and was wondering if anyone out there in Internet-land has: (1) used ovalbumin as a model glycoprotein for receptor binding studies. I've biotinylated it using iodiacetyl-biotin, fed it to cells at 4 degrees C, and then gone in with fluorescent avidin to detect binding. However, I'm not sure if it will actually bind. Anyone know? (2) used RAW 264.7 cells (a macrophage-like cell line) for studying the macrophage mannose receptor. I've got some more tests to do, but I was wondering if anybody can help me out. Thanks in advance! -Mara From owner-proteins@net.bio.net Fri Feb 09 22:00:00 1996 Newsgroups: bionet.cellbiol,bionet.molbio.methds-reagnts,bionet.molbio.proteins Path: biosci!news.Stanford.EDU!nntp-hub2.barrnet.net!newsfeed.internetmci.com!swrinde!sgigate.sgi.com!sdd.hp.com!col.hp.com!csn!ub!hood.cc.rochester.edu!uhura.cc.rochester.edu!ttha From: ttha@uhura.cc.rochester.edu (Tom Thatcher) Subject: Re: Silver staining problem Message-ID: <1996Feb11.034525.25089@galileo.cc.rochester.edu> Sender: news@galileo.cc.rochester.edu Nntp-Posting-Host: uhura.cc.rochester.edu Organization: University of Rochester - Rochester, New York References: <4fj93s$80q@sage.cc.purdue.edu> Date: Sun, 11 Feb 96 03:45:25 GMT Lines: 52 Xref: biosci bionet.cellbiol:4013 bionet.molbio.methds-reagnts:40134 bionet.molbio.proteins:7016 In <4fj93s$80q@sage.cc.purdue.edu> hamburg@sage.cc.purdue.edu (Eric Sarnighausen) writes: >I am analyzing patterns of cell wall proteins by two dimensional >electrophoresis. Unfortunately, the most prominent protein of 23 kDa >(that I am extremely interested in) is not silver stained at all (using >the acidic method). I have not had that specific problem but here is my favorite silver stain. Sometimes a different technique gives different results, and everyone I've shared this with goes bonkers over it. Silver Staining of SDS Protein Gels Reference: Lischwe and Ochs, 1982. Anal. Biochem. 127:453-457. All operations should be carried out in glass trays. If the gel is touched, fingerprints will appear when it is developed. 1. Fix the gel in 50% ethanol, 10% acetic acid at least 1 hour, with gentle shaking. 2. Wash the gel in 10% ethanol, 5% acetic acid, 2 times for 30 minutes each, with gentle shaking. 3. Incubate the gel in 100 ml 0.02% AgNO3 (0.2 g AgNO3 per 100 ml H2O) for 1 to 2 hours, with gentle shaking 4. Aspirate or pour the silver solution into an appropriate waste container. Rinse the gel for a few seconds in distilled H2O. 5. The developer is 4.5 g NaOH, 13 mg NaBH3 (sodium borohydride), and 1 ml formaldehyde (commercial stock solution, 37% formaldehyde) in 150 ml H2O. Develop the gel. Pour a few ml developer into the tray with the gel. A black precipitate will form. Quickly aspirate away this precipitate, then add the rest of the developer. Optimum developing time for a 0.75 mm thick gel is 4 to 5 minutes; slightly longer for thicker gels. 6. Incubate the gel in 100 ml 0.75% Na2CO3 for three changes of 30 minutes each. This enhances and stabilizes the silver stain. The complete treatment is not absolutely necessary, but the gel should be treated at least once. 7. Soak the gel in 3% acetic acid to stop further development. The gel will eventually fade (in a few days) unless it is dried down. -- Tom Thatcher | You can give a PC to a Homo habilis, University of Rochester Cancer Center | and he'll use it, but he'll use it ttha@uhura.cc.rochester.edu | to crack nuts. From owner-proteins@net.bio.net Fri Feb 09 22:00:00 1996 Path: biosci!bloom-beacon.mit.edu!newsxfer2.itd.umich.edu!newsfeed.internetmci.com!vixen.cso.uiuc.edu!howland.reston.ans.net!usenet.ins.cwru.edu!slider.bme.ri.ccf.org!kira.cc.uakron.edu!odin.oar.net!malgudi.oar.net!news.cas.org!usenet From: rag55@cas.org (Roger Granet) Newsgroups: bionet.molbio.proteins Subject: Re: HISTONE ACETYLASES/DEACETYLASES Date: 9 Feb 1996 17:57:21 GMT Organization: Chemical Abstracts Service Lines: 58 Message-ID: <4fg1u1$2qv@srv13s4.cas.org> References: <4f8kas$sma@nntp.Stanford.EDU> Reply-To: rag55@cas.org NNTP-Posting-Host: rag55mws.cas.org Keywords: online info., histone acetylase/deacetylase gene cloning, CAS In article <4f8kas$sma@nntp.Stanford.EDU>, Ignacio Marin writes: > Is there any information available about clones histone > acetylases/deacetylases in any organism?. Any information will be > welcome. > > Ignacio Marin. Dpt. Biological Sciences. Stanford University. Ignacio, Hi. From a quick search of the CAplus file produced by the organization I work at, Chemical Abstracts Service, I found 158 abstracts relating to histone acetylases or deacetylases. Of these, only two related to their cloning, however. They were: o TI Identification of a gene encoding a yeast histone H4 acetyltransferase AU Kleff, Susanne; Andrulis, Erik D.; Anderson, Carl W.; Sternglanz, Rolf CS Dep. Biochem. Cell Biol., State Univ. New York, Stony Brook, N. Y., NY, 11794, USA SO J. Biol. Chem. (1995), 270(42), 24674-7 o TI The predicted molecular structure suggests that CTF/NF-I may function as a histone acetylase AU Oikarinen, Jouko; Mannermaa, Riitta Maaria CS Biocent., Univ. Oulu, Oulu, SF-90220, Finland SO FEBS Lett. (1990), 273(1-2), 11-14 I hope these will be of some use. A lot of people think that Chemical Abstracts has only purely chemical information, but, a third of the 680,000 or so abstracts (journals, patents, conference proceedings, etc.) we put out every year come from our bio-related areas and include molecular genetics abstracts like the above ones. Good luck! Roger Roger Granet Phone: (614) 447-3600, ext. 2346 Associate Scientific Information Analyst Internet: rgranet@cas.org Biochemistry Department Chemical Abstracts Service P.O. box 3012 Columbus, OH 43210 CAS Customer Service: 1-800-753-4227 or by email to: help@cas.org STN Help Desk: 1-800-848-6533 (in North America) or (+49)7247/808-555 (in Europe) or (+81)3-5214-8413 (in Asia) Web site at: http://info.cas.org/welcome.html The views given here are mine only and do not necessarily represent those of anyone else, Chemical Abstracts Service, or the American Chemical Society. From owner-proteins@net.bio.net Fri Feb 09 22:00:00 1996 Path: biosci!bcm.tmc.edu!newshost.convex.com!news.duke.edu!news.mathworks.com!newsfeed.internetmci.com!uwm.edu!math.ohio-state.edu!usc!newshub.cts.com!meador.cts.com!user From: meador@cts.com (Jim) Newsgroups: bionet.molbio.methds-reagnts,bionet.cellbiol,bionet.microbiology,bionet.molbio.proteins,sci.bio.microbiology Subject: Re: bacterial housekeeping genes Date: Sat, 10 Feb 1996 09:27:28 -0800 Organization: CTS Network Services Lines: 45 Message-ID: Referenc